SUBSTITUTED HETEROCYCLES AS c-MYC TARGETING AGENTS

ABSTRACT

Disclosed are substituted heterocycle compounds including substituted pyrazoles, substituted pyrimidines, and substitute triazoles. The substituted heterocycles disclosed herein are shown to be useful in inhibiting c-MYC and may be utilized as therapeutics for treating cancer and cell proliferative disorders.

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

This application claims the benefit of priority under 35 U.S.C. § 119(e)to U.S. Provisional Application No. 62/862,304, filed on Jun. 17, 2019,the content of which is incorporated herein by reference in itsentirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under grant numberCA123484 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND

The field of the invention relates to substituted heterocycles as c-MYCtargeting agents. In particular, the field of the invention relates tosubstituted pyrazoles, pyrimidines, or triazoles as c-MYC targetingagents for the treatment of cell proliferation diseases and disorderssuch as cancer.

The c-MYC oncogene is de-regulated and plays a causal role in a majorityof human cancer and c-MYC inhibition profoundly affects tumor growth orsurvival in multiple models. MYC is the most common oncogene involved inhuman cancers and is overexpressed in up to half of all cancers.Therefore, developing c-MYC inhibitors is among the most attractivepotential anti-cancer strategies. Unfortunately, due to the difficultyin targeting transcription factors with small molecules, c-MYC iscurrently regarded as “undruggable.” Here, we disclose a new approach totargeting c-MYC and have developed a series of new small moleculeinhibitors. These compounds selectively target c-MYC-driven cellproliferation and interfere with binding of c-MYC to DNA.

SUMMARY

Disclosed are substituted heterocycles which may be utilized as c-MYCtargeting agents. The substituted heterocycles may include substitutedpyrazoles, substituted pyrimidines, and substituted triazoles. Thedisclosed heterocycles may be used in pharmaceutical compositions andmethods for treating cell proliferative disorders such as cancer.

The disclosed substituted heterocycles may include substituted pyrazoleshaving a formula I

wherein

-   R¹ is hydrogen or halo, or R¹ is alkyl, alkoxy, hydroxyl, aryl    (e.g., phenyl), alkylaryl (e.g., benzyl), alkyldiaryl (e.g.,    methyldiphenyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl,    pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,    pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,    furan-2-yl, furan-3-yl, imidazo[1,2-A]pyridine such as    imidazo[1,2-A]pyridin-4-yl, imidazo[1,2-A]pyridin-5-yl,    imidazo[1,2-A]pyridin-6-yl, imidazo[1,2-A]pyridin-7-yl,    imidazo[1,2-A]pyridin-8-yl, or imidazo[1,2-A]pyridin-9-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl such as N-piperidinyl, piperidin-2-yl, piperidin-3-yl,    or piperidin-4-yl; piperazinyl such as N-piperazinyl; morpholinyl    such as N-morpholinyl; tetrahydropyranyl such as    tetrahydropyran-4-yl), optionally R¹ is substituted at one or more    positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and    alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   n is 0 or 1;-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   Y is C, CH, C-halo (e.g., C—F, C—Br, C—Cl, or C—I), C-haloalkyl    (e.g., C—CF₃), or N;-   Z is C, CH, C-halo (e.g., C—F, C—Br, C—Cl, or C—I), C-haloalkyl    (e.g., C—CF₃), or N;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R³ is hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆    alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), aryl (e.g., phenyl), alkylaryl (e.g., benzyl),    hydroxyl, halo, carboxyamido, hydrazonyl, carbonyl, carboxyl, or    alkoxycarbonyl;-   R⁴ is present or absent and when present R⁴ is hydrogen, amino,    alkyl, or R⁴ is aryl (e.g., phenyl), or alkylaryl (e.g., benzyl); R⁴    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy);-   W is C or N;-   R⁵ is present or absent and when present R⁵ is hydrogen, alkyl    (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl (e.g.,    trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl, or    halo;-   R⁶ is present or absent and when present R⁶ is hydrogen, amino,    alkyl, or R⁶ is aryl (e.g., phenyl) or alkylaryl (e.g., benzyl); R⁶    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy), or R⁶ and R⁵ together form a ring    structure having a formula

-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, pyrimidinyl (e.g., pyrimidin-2-yl,    pyrimidin-4-yl, pyrimidin-5-yl, or pyrimidin-6-yl),    1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl, morpholinyl), optionally R⁷ is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,    hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo; and-   with the proviso that at least one of R⁴ and R⁶ is absent;-   optionally with the proviso that if R⁵ is hydrogen, then p is 1 and    m is 1;-   optionally with the proviso that if R¹(Alk¹)_(n)(X)_(p)(Alk²)_(q)-    is hydrogen, hydroxyl, or alkyl, and R⁵ is hydroxyl, then m is 1, or    at least one of R² and R³ is not hydrogen; and-   optionally with the proviso that no more than 2 of W, Y, and Z are    N.

In the disclosed formula I, Pyr is a pyrazole ring having twonon-adjacent double bonds, for example, where the substituted pyrazoleshave a formula I(i) or I(ii):

Specifically, the substituted pyrazoles may have a formula Ia(i),Ia(ii), Ib(i), Ib(ii), Ic(i), or Ic(ii):

The disclosed compounds may exhibit one or more biological activities.The disclosed compounds may inhibit binding of the MYC/Max complex toDNA (e.g., in a DNA gel shifting assay). The disclosed compounds may notproduce significant DNA damage (e.g., in an rH2AX staining assay at aconcentration greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.1 μM,1.0 μM, 10 μM, 100 μM, or higher). The disclosed compounds may inhibitthe growth of cells that express c-MYC (preferably by at a concentrationof less than about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM,0.005 μM, 0.001 μM, or less). The disclosed compounds may not inhibitthe growth of cells that do not express c-MYC (preferably at aconcentration of greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM,0.1 μM, 1.0 μM, 10 μM, and 100 μM or higher).

Also disclosed are pharmaceutical compositions comprising the disclosedcompounds and a suitable pharmaceutical carrier, excipient, or diluent.The disclosed pharmaceutical compositions may comprise an effectiveamount of the compound for inhibiting the growth of cancer cells whenadministered to a subject in need thereof.

Also disclosed are methods for treating cell proliferation diseases anddisorders such as cancer. The methods may include administering thedisclosed compounds or pharmaceutical compositions comprising thedisclosed compounds to a subject in need thereof, for example, to asubject having cancer. The disclosed compounds or pharmaceuticalcompositions comprising the disclosed compounds may be administered withadditional therapeutic agents, optionally in combination, in order totreat cell proliferative diseases and disorders. Cell proliferativediseases and disorders treated by the disclosed methods may include, butare not limited to, cancers selected from the group consisting ofmultiple myeloma, leukemia, non-small cell lung cancer, colon cancer,cancer of the central nervous system, melanoma, ovarian cancer, renalcancer, prostate cancer, and breast cancer.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Scheme for in silico screen to identify c-MYC inhibitors.

FIG. 2. Relative MYC E-box luciferase inhibitory activity of 32compounds.

FIG. 3. Growth inhibition by selected hit compound on MYC WT and KOfibroblasts.

FIG. 4. Cell viability after treatment with Min9 (NUCC-0176234).

FIG. 5. Electrophoretic mobility shift assay (EMSA) in the presence of200 μM test compounds.

FIG. 6. Relative values of DNA bound for test compounds at 200 μM.

FIG. 7. Relative MYC/Max DNA binding versus concentration of compound.

FIG. 8. rH2AX assay for DNA damage.

FIG. 9. In vitro metabolism of NUCC-176242 versus NUCC-176248.

FIG. 10. Pharmacokinetic study of NUCC-176242 in mice after IV dosing at5 mg/kg.

FIG. 11. Myc inhibitor 361 specifically inhibits proliferation of Mycdependent cell line PC3, but not Myc independent cell line PC12.

FIG. 12. Myc inhibitor 361 inhibits c-Myc transcription activity as anearly event before affecting cell viability.

FIG. 13. Myc inhibitor 361 inhibits c-Myc transcription activity as anearly event before affecting cell viability.

FIG. 14. MYC inhibitor Min9-S1 impairs MYC transcriptional program.

FIG. 15. Myc inhibitor 361 reduces tumor volume in a mouse prostatecancer allograft model.

FIG. 16. Myc inhibitor 361 combination with Immunotherapy significantlyblocks tumor progression.

FIG. 17. Myc inhibitor 975 and 031 shows significant synergic anti-tumoreffect with Ara-C in a AML mice xenograft model.

FIG. 18. NCI60 profiling of compound 201195 at a single concentration of10 uM.

FIG. 19. NCI60 profiling of compound 200975 at a single concentration of10 uM.

FIG. 20. NCI60 profiling of compound 196361 at a single concentration of10 uM.

DETAILED DESCRIPTION

The present invention is described herein using several definitions, asset forth below and throughout the application.

Unless otherwise specified or indicated by context, the terms “a”, “an”,and “the” mean “one or more.” For example, “a compound” and “asubstituent” should be interpreted to mean “one or more compounds” and“one or more substituents,” respectively.

As used herein, “about,” “approximately,” “substantially,” and“significantly” will be understood by persons of ordinary skill in theart and will vary to some extent on the context in which they are used.If there are uses of these terms which are not clear to persons ofordinary skill in the art given the context in which they are used,“about” and “approximately” will mean plus or minus ≤10% of theparticular term and “substantially” and “significantly” will mean plusor minus >10% of the particular term.

As used herein, the terms “include” and “including” have the samemeaning as the terms “comprise” and “comprising” in that these latterterms are “open” transitional terms that do not limit claims only to therecited elements succeeding these transitional terms. The term“consisting of,” while encompassed by the term “comprising,” should beinterpreted as a “closed” transitional term that limits claims only tothe recited elements succeeding this transitional term. The term“consisting essentially of,” while encompassed by the term “comprising,”should be interpreted as a “partially closed” transitional term whichpermits additional elements succeeding this transitional term, but onlyif those additional elements do not materially affect the basic andnovel characteristics of the claim.

As used herein, a “subject” may be interchangeable with “patient” or“individual” and means an animal, which may be a human or non-humananimal, in need of treatment.

A “subject in need of treatment” may include a subject having a disease,disorder, or condition that is responsive to therapy with a substitutedheterocycle such as the presently disclosed substituted pyrazoles,substituted pyrimidines, and substituted triazoles. For example, a“subject in need of treatment” may include a subject having a cellproliferative disease, disorder, or condition such as cancer (e.g.,cancers such as multiple myeloma, leukemia, non-small cell lung cancer,colon cancer, cancer of the central nervous system, melanoma, ovariancancer, renal cancer, prostate cancer, and breast cancer). A “subject inneed of treatment” may include a subject having a cell proliferativedisease, disorder, or condition such as cancer that is associated withc-MYC activity and/or that may be treated by administering an effectiveamount of an agent that modulates c-MYC activity.

As used herein, the phrase “effective amount” shall mean that drugdosage that provides the specific pharmacological response for which thedrug is administered in a significant number of subject in need of suchtreatment. An effective amount of a drug that is administered to aparticular subject in a particular instance will not always be effectivein treating the conditions/diseases described herein, even though suchdosage is deemed to be a therapeutically effective amount by those ofskill in the art.

As used herein, the term “modulate” means decreasing or inhibitingactivity and/or increasing or augmenting activity. For example,modulating c-MYC activity may mean increasing or augmenting c-MYCactivity and/or decreasing or inhibiting c-MYC activity. The compoundsdisclosed herein may be administered to modulate c-MYC activity.

Chemical Entities

New chemical entities and uses for chemical entities are disclosedherein. The chemical entities may be described using terminology knownin the art and further discussed below.

As used herein, an asterisk “*” or a plus sign “+” may be used todesignate the point of attachment for any radical group or substituentgroup.

The term “alkyl” as contemplated herein includes a straight-chain orbranched alkyl radical in all of its isomeric forms, such as a straightor branched group of 1-12, 1-10, or 1-6 carbon atoms, referred to hereinas C1-C12 alkyl, C1-C10-alkyl, and C1-C6-alkyl, respectively.

The term “alkylene” refers to a diradical of straight-chain or branchedalkyl group (i.e., a diradical of straight-chain or branched C₁-C6 alkylgroup). Exemplary alkylene groups include, but are not limited to —CH₂—,—CH₂CH₂—, —CH₂CH₂CH₂—, —CH(CH₃)CH₂—, —CH₂CH(CH₃)CH₂—, —CH(CH₂CH₃)CH₂—,and the like.

The term “haloalkyl” refers to an alkyl group that is substituted withat least one halogen. For example, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CF₂CF₃,and the like.

The term “heteroalkyl” as used herein refers to an “alkyl” group inwhich at least one carbon atom has been replaced with a heteroatom(e.g., an O, N, or S atom). One type of heteroalkyl group is an “alkoxy”group.

The term “alkenyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon double bond, suchas a straight or branched group of 2-12, 2-10, or 2-6 carbon atoms,referred to herein as C2-C12-alkenyl, C2-C10-alkenyl, and C2-C6-alkenyl,respectively.

The term “alkynyl” as used herein refers to an unsaturated straight orbranched hydrocarbon having at least one carbon-carbon triple bond, suchas a straight or branched group of 2-12, 2-10, or 2-6 carbon atoms,referred to herein as C2-C12-alkynyl, C2-C10-alkynyl, and C2-C6-alkynyl,respectively.

The term “cycloalkyl” refers to a monovalent saturated cyclic, bicyclic,or bridged cyclic (e.g., adamantyl) hydrocarbon group of 3-12, 3-8, 4-8,or 4-6 carbons, referred to herein, e.g., as “C4-8-cycloalkyl,” derivedfrom a cycloalkane. Unless specified otherwise, cycloalkyl groups areoptionally substituted at one or more ring positions with, for example,alkanoyl, alkoxy, alkyl, haloalkyl, alkenyl, alkynyl, amido orcarboxyamido, amidino, amino, aryl, arylalkyl, azido, carbamate,carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halo,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro,phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido,sulfonyl or thiocarbonyl. In certain embodiments, the cycloalkyl groupis not substituted, i.e., it is unsubstituted.

The term “cycloheteroalkyl” refers to a monovalent saturated cyclic,bicyclic, or bridged cyclic hydrocarbon group of 3-12, 3-8, 4-8, or 4-6carbons in which at least one carbon of the cycloalkane is replaced witha heteroatom such as, for example, N, O, and/or S.

The term “cycloalkylene” refers to a cycloalkyl group that isunsaturated at one or more ring bonds.

The term “partially unsaturated carbocyclyl” refers to a monovalentcyclic hydrocarbon that contains at least one double bond between ringatoms where at least one ring of the carbocyclyl is not aromatic. Thepartially unsaturated carbocyclyl may be characterized according to thenumber oring carbon atoms. For example, the partially unsaturatedcarbocyclyl may contain 5-14, 5-12, 5-8, or 5-6 ring carbon atoms, andaccordingly be referred to as a 5-14, 5-12, 5-8, or 5-6 memberedpartially unsaturated carbocyclyl, respectively. The partiallyunsaturated carbocyclyl may be in the form of a monocyclic carbocycle,bicyclic carbocycle, tricyclic carbocycle, bridged carbocycle,spirocyclic carbocycle, or other carbocyclic ring system. Exemplarypartially unsaturated carbocyclyl groups include cycloalkenyl groups andbicyclic carbocyclyl groups that are partially unsaturated. Unlessspecified otherwise, partially unsaturated carbocyclyl groups areoptionally substituted at one or more ring positions with, for example,alkanoyl, alkoxy, alkyl, haloalkyl, alkenyl, alkynyl, amido orcarboxyamido, amidino, amino, aryl, arylalkyl, azido, carbamate,carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen,haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro,phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamido,sulfonyl or thiocarbonyl. In certain embodiments, the partiallyunsaturated carbocyclyl is not substituted, i.e., it is unsubstituted.

The term “aryl” is art-recognized and refers to a carbocyclic aromaticgroup. Representative aryl groups include phenyl, naphthyl, anthracenyl,and the like. The term “aryl” includes polycyclic ring systems havingtwo or more carbocyclic rings in which two or more carbons are common totwo adjoining rings (the rings are “fused rings”) wherein at least oneof the rings is aromatic and, e.g., the other ring(s) may becycloalkyls, cycloalkenyls, cycloalkynyls, and/or aryls. Unlessspecified otherwise, the aromatic ring may be substituted at one or morering positions with, for example, halogen, azide, alkyl, aralkyl,alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro,sulfhydryl, imino, amido or carboxyamido, carboxylic acid, —C(O)alkyl,—CO₂alkyl, carbonyl, carboxyl, alkylthio, sulfonyl, sulfonamido,sulfonamide, ketone, aldehyde, ester, heterocyclyl, aryl or heteroarylmoieties, —CF₃, —CN, or the like. In certain embodiments, the aromaticring is substituted at one or more ring positions with halogen, alkyl,hydroxyl, or alkoxyl. In certain other embodiments, the aromatic ring isnot substituted, i.e., it is unsubstituted. In certain embodiments, thearyl group is a 6-10 membered ring structure.

The terms “heterocyclyl” and “heterocyclic group” are art-recognized andrefer to saturated, partially unsaturated, or aromatic 3- to 10-memberedring structures, alternatively 3- to 7-membered rings, whose ringstructures include one to four heteroatoms, such as nitrogen, oxygen,and sulfur. The number of ring atoms in the heterocyclyl group can bespecified using 5 Cx-Cx nomenclature where x is an integer specifyingthe number of ring atoms. For example, a C3-C7 heterocyclyl group refersto a saturated or partially unsaturated 3- to 7-membered ring structurecontaining one to four heteroatoms, such as nitrogen, oxygen, andsulfur. The designation “C3-C7” indicates that the heterocyclic ringcontains a total of from 3 to 7 ring atoms, inclusive of any heteroatomsthat occupy a ring atom position.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines (e.g., mono-substituted amines ordi-substituted amines), wherein substituents may include, for example,alkyl, cycloalkyl, heterocyclyl, alkenyl, and aryl.

The terms “alkoxy” or “alkoxyl” are art-recognized and refer to an alkylgroup, as defined above, having an oxygen radical attached thereto.Representative alkoxy groups include methoxy, ethoxy, tert-butoxy andthe like.

An “ether” is two hydrocarbons covalently linked by an oxygen.Accordingly, the substituent of an alkyl that renders that alkyl anether is or resembles an alkoxyl, such as may be represented by one of—O-alkyl, —O-alkenyl, —O-alkynyl, and the like.

The term “carbonyl” as used herein refers to the radical —C(O)—.

The term “oxo” refers to a divalent oxygen atom —O—.

The term “carboxamido” as used herein refers to the radical —C(O)NRR′,where R and R′ may be the same or different. R and R′, for example, maybe independently hydrogen, alkyl, aryl, arylalkyl, cycloalkyl, formyl,haloalkyl, heteroaryl, or heterocyclyl.

The term “carboxy” as used herein refers to the radical —COOH or itscorresponding salts, e.g. —COONa, etc.

The term “amide” or “amido” or “amidyl” as used herein refers to aradical of the form —R¹C(O)N(R²)—, —R¹C(O)N(R²)R³—, —C(O)NR²R³, or—C(O)NH₂, wherein R¹, R² and R³, for example, are each independentlyhydrogen, alkyl, alkoxy, alkenyl, alkynyl, amide, amino, aryl,arylalkyl, carbamate, cycloalkyl, ester, ether, formyl, halogen,haloalkyl, heteroaryl, heterocyclyl, hydrogen, hydroxyl, ketone, ornitro.

The compounds of the disclosure may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asgeometric isomers, enantiomers or diastereomers. The term“stereoisomers” when used herein consist of all geometric isomers,enantiomers or diastereomers. These compounds may be designated by thesymbols “R” or “S,” or “+” or “−” depending on the configuration ofsubstituents around the stereogenic carbon atom and or the opticalrotation observed. The present invention encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers includeenantiomers and diastereomers. Mixtures of enantiomers or diastereomersmay be designated (±)″ in nomenclature, but the skilled artisan willrecognize that a structure may denote a chiral center implicitly. It isunderstood that graphical depictions of chemical structures, e.g.,generic chemical structures, encompass all stereoisomeric forms of thespecified compounds, unless indicated otherwise. Also contemplatedherein are compositions comprising, consisting essentially of, orconsisting of an enantiopure compound, which composition may comprise,consist essential of, or consist of at least about 50%, 60%, 70%, 80%,90%, 95%, 96%, 97%, 98%, 99%, or 100% of a single enantiomer of a givencompound (e.g., at least about 99% of an R enantiomer of a givencompound).

Substituted Heterocycles and Uses Thereof for Inhibiting the BiologicalActivity of C-MYC

Disclosed herein are substituted heterocycles. The disclosedheterocycles have been shown to inhibit the biological activity ofc-MYC. The disclosed substituted heterocycles may include substitutedpyrazoles, substituted pyrimidines, and substituted triazoles.

In some embodiments, the disclosed substituted heterocycles may includesubstituted pyrazoles having a formula I:

wherein

-   R¹ is hydrogen or halo, or R¹ is alkyl, alkoxy, hydroxyl, aryl    (e.g., phenyl), alkylaryl (e.g., a benzyl), alkyldiaryl (e.g.,    methyldiphenyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl,    pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,    pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,    furan-2-yl, furan-3-yl, imidazo[1,2-A]pyridine such as    imidazo[1,2-A]pyridin-4-yl, imidazo[1,2-A]pyridin-5-yl,    imidazo[1,2-A]pyridin-6-yl, imidazo[1,2-A]pyridin-7-yl,    imidazo[1,2-A]pyridin-8-yl, or imidazo[1,2-A]pyridin-9-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl such as N-piperidinyl, piperidin-2-yl, piperidin-3-yl,    or piperidin-4-yl; piperazinyl such as N-piperazinyl; morpholinyl    such as N-morpholinyl; tetrahydropyranyl such as    tetrahydropyran-4-yl), optionally R¹ is substituted at one or more    positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and    alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   n is 0 or 1;-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   Y is C, CH, C-halo (e.g., C—F, C—Br, C—Cl, or C—I), C-haloalkyl    (e.g., C—CF₃), or N;-   Z is C, CH, C-halo (e.g., C—F, C—Br, C—Cl, or C—I), C-haloalkyl    (e.g., C—CF₃), or N;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R³ is hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆    alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), aryl (e.g., phenyl), alkylaryl (e.g., benzyl),    hydroxyl, halo, carboxyamido, hydrazonyl, carbonyl, carboxyl, or    alkoxycarbonyl;-   R⁴ is present or absent and when present R⁴ is hydrogen, amino,    alkyl, or R⁴ is aryl (e.g., phenyl), or alkylaryl (e.g., benzyl); R⁴    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy);-   W is C or N;-   R⁵ is present or absent and when present R⁵ is hydrogen, alkyl    (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl (e.g.,    trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl, or    halo;-   R⁶ is present or absent and when present R⁶ is hydrogen, amino,    alkyl, or R⁶ is aryl (e.g., phenyl) or alkylaryl (e.g., benzyl); R⁶    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy), or R⁶ and R⁵ together form a ring    structure having a formula

-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, pyrimidinyl (e.g., pyrimidin-2-yl,    pyrimidin-4-yl, pyrimidin-5-yl, or pyrimidin-6-yl),    1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl, morpholinyl), optionally R⁷ is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,    hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo; and-   optionally with the proviso that at least one of R⁴ and R⁶ is    hydrogen;-   optionally with the proviso that if R⁵ is hydrogen, then p is 1 and    m is 1;-   optionally with the proviso that if R¹(Alk¹)_(n)(X)_(p)(Alk²)_(q)-    is hydrogen, hydroxyl, or alkyl, and R⁵ is hydroxyl, then m is 1, or    at least one of R² and R³ is not hydrogen; and-   optionally with the proviso that no more than 2 of W, Y, and Z are    N.

In some embodiments of these disclosed substituted pyrazoles, at leastone of R² and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR² and R⁷ optionally are substituted at one or more positions with oneor more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy),hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl. In some embodiments of the disclosed compounds, m is 0and R² is hydrogen, or R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, R² is aryl(e.g., phenyl), alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl,furan-3-yl), cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,piperidinyl, morpholinyl), and R² optionally is substituted at one ormore positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy(e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy(e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl; and R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, m is 0 andR² is hydrogen; and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR⁷ optionally is substituted at one or more positions with one or moreof alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl(e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl.

In the disclosed formula I, Pyr is a pyrazole ring having twonon-adjacent double bonds, for example, where the substituted pyrazoleshave a formula I(i) or I(ii):

Specifically, the substituted pyrazoles may have a formula Ia(i),Ia(ii), Ib(i), Ib(ii), Ic(i), or Ic(ii):

In some embodiments, the disclosed substituted heterocycles may includesubstituted pyrazoles having a formula II:

wherein

-   R¹ is hydrogen or halo, or R¹ is alkyl, alkoxy, hydroxyl, aryl    (e.g., phenyl), alkylaryl (e.g., benzyl), alkyldiaryl (e.g.,    methyldiphenyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl,    pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,    pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,    furan-2-yl, furan-3-yl, imidazo[1,2-A]pyridine such as    imidazo[1,2-A]pyridin-4-yl, imidazo[1,2-A]pyridin-5-yl,    imidazo[1,2-A]pyridin-6-yl, imidazo[1,2-A]pyridin-7-yl,    imidazo[1,2-A]pyridin-8-yl, or imidazo[1,2-A]pyridin-9-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl such as N-piperidinyl, piperidin-2-yl, piperidin-3-yl,    or piperidin-4-yl; piperazinyl such as N-piperazinyl; morpholinyl    such as N-morpholinyl; tetrahydropyranyl such as    tetrahydropyran-4-yl), optionally R¹ is substituted at one or more    positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and    alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   Y is CH or N;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R³ is hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆    alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), aryl (e.g., phenyl), alkylaryl (e.g., benzyl),    hydroxyl, halo, carboxyamido, hydrazonyl, carbonyl, carboxyl, or    alkoxycarbonyl;-   R⁴ is present or absent and when present R⁴ is hydrogen, amino,    alkyl, or R⁴ is aryl (e.g., phenyl), or alkylaryl (e.g., benzyl); R⁴    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy);-   R⁶ is present or absent and when present R⁶ is hydrogen, amino,    alkyl, or R⁶ is aryl (e.g., phenyl) or alkylaryl (e.g., benzyl); R⁶    optionally is substituted at one or more positions with one or more    of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl    (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), aryl    (e.g., phenyl), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, alkoxycarbonyl, aryloxy (e.g., phenoxy), and    alkylaryloxy (e.g., benzyloxy), or R⁶ and R⁵ together form a ring    structure having a formula

-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, pyrimidinyl (e.g., pyrimidin-2-yl,    pyrimidin-4-yl, pyrimidin-5-yl, or pyrimidin-6-yl),    1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl, morpholinyl), optionally R⁷ is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,    hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo; and-   with the proviso that at least one of R⁴ and R⁶ is absent; and-   optionally with the proviso that if R⁵ is hydrogen, then p is 1 and    m is 1.

In some embodiments of these disclosed substituted pyrazoles, at leastone of R² and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR² and R⁷ optionally are substituted at one or more positions with oneor more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy),hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl. In some embodiments of the disclosed compounds, m is 0and R² is hydrogen, or R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, R² is aryl(e.g., phenyl), alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl,furan-3-yl), cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,piperidinyl, morpholinyl), and R² optionally is substituted at one ormore positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy(e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy(e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl; and R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, m is 0 andR² is hydrogen; and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR⁷ optionally is substituted at one or more positions with one or moreof alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl(e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl.

Specifically, the substituted pyrazoles may have a formula IIa:

In some embodiments, the disclosed substituted heterocycles may includesubstituted pyrimidines having a formula III:

wherein:

-   R¹ is hydrogen or halo, or R¹ is alkyl, alkoxy, hydroxyl, aryl    (e.g., phenyl), alkylaryl (e.g., a benzyl), alkyldiaryl (e.g.,    methyldiphenyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl,    pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,    pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,    furan-2-yl, furan-3-yl, imidazo[1,2-A]pyridine such as    imidazo[1,2-A]pyridin-4-yl, imidazo[1,2-A]pyridin-5-yl,    imidazo[1,2-A]pyridin-6-yl, imidazo[1,2-A]pyridin-7-yl,    imidazo[1,2-A]pyridin-8-yl, or imidazo[1,2-A]pyridin-9-yl),    cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,    piperidinyl such as N-piperidinyl, piperidin-2-yl, piperidin-3-yl,    or piperidin-4-yl; piperazinyl such as N-piperazinyl; morpholinyl    such as N-morpholinyl; tetrahydropyranyl such as    tetrahydropyran-4-yl), optionally R¹ is substituted at one or more    positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and    alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   n is 0 or 1;-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R⁴ is hydrogen, amino, alkyl, or R⁴ is aryl (e.g., phenyl), or    alkylaryl (e.g., benzyl); R⁴ optionally is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, alkoxycarbonyl,    aryloxy (e.g., phenoxy), and alkylaryloxy (e.g., benzyloxy);-   R⁵ is hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆    alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, or halo;-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ alkyl, aryl (e.g., phenyl), alkylaryl    (e.g., benzyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl,    pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,    pyrazol-4-yl, pyrimidinyl (e.g., pyrimidin-2-yl, pyrimidin-4-yl,    pyrimidin-5-yl, or pyrimidin-6-yl), 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R⁷ is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo;-   optionally with the proviso that if R⁵ is hydrogen, then p is 1 and    m is 1;-   optionally with the proviso that if R¹(Alk¹)_(n)(X)_(p)(Alk²)_(q)-    is hydrogen, hydroxyl, or alkyl, and R⁵ is hydroxyl, then m is 1, or    R² is not hydrogen.

In some embodiments of these disclosed substituted pyrimidines, at leastone of R² and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR² and R⁷ optionally are substituted at one or more positions with oneor more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy),hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl. In some embodiments of the disclosed compounds, m is 0and R² is hydrogen, or R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrimidines, R² isaryl (e.g., phenyl), alkylaryl (e.g., benzyl), heteroaryl (e.g.,N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,furan-2-yl, furan-3-yl), cycloalkyl (e.g., cyclohexyl), orcycloheteroalkyl (e.g., piperidinyl, morpholinyl), and R² optionally issubstituted at one or more positions with one or more of alkyl (e.g.,C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl (e.g.,trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl, halo,cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;and R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrimidines, m is 0and R² is hydrogen; and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g.,benzyl), heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl,pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl,1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl),cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl,morpholinyl), and R⁷ optionally is substituted at one or more positionswith one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, and alkoxycarbonyl.

Specifically, the substituted pyrimidines may have a formula IIIa orIIb:

wherein:

-   R⁴ is hydrogen, amino, alkyl, or R⁴ is aryl (e.g., phenyl), or    alkylaryl (e.g., benzyl); R⁴ optionally is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, alkoxycarbonyl,    aryloxy (e.g., phenoxy), and alkylaryloxy (e.g., benzyloxy)

In some embodiments, the disclosed substituted heterocycles may includesubstitute pyrazoles having a formula IV:

wherein:

-   R¹ is hydrogen, or R¹ is aryl (e.g., phenyl), alkylaryl (e.g.,    benzyl), alkyldiaryl (e.g., methyldiphenyl), heteroaryl (e.g.,    N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R¹ is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo, cyano,    carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   n is 0 or 1;-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   Y is CH or N;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g. benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R⁴ is hydrogen, amino, alkyl, or R⁴ is aryl (e.g., phenyl) or    alkylaryl (e.g., benzyl); R⁴ optionally is substituted at one or    more positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy    (e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy    (e.g., trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo,    cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, alkoxycarbonyl,    aryloxy (e.g., phenoxy), and alkylaryloxy (e.g., benzyloxy);-   R⁵ is present or absent and when present R⁵ is hydrogen, alkyl    (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl (e.g.,    trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl, or    halo;-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R⁷ is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl; and-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo.

In some embodiments of these disclosed substituted pyrazoles, at leastone of R² and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR² and R⁷ optionally are substituted at one or more positions with oneor more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy),hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl. In some embodiments of the disclosed compounds, m is 0and R² is hydrogen, or R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, R² is aryl(e.g., phenyl), alkylaryl (e.g., a benzyl), heteroaryl (e.g.,N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl,furan-2-yl, furan-3-yl), cycloalkyl (e.g., cyclohexyl), orcycloheteroalkyl (e.g., piperidinyl, morpholinyl), and R² optionally issubstituted at one or more positions with one or more of alkyl (e.g.,C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl (e.g.,trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl, halo,cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;and R⁷ is hydrogen.

In some embodiments of these disclosed substituted pyrazoles, m is 0 andR² is hydrogen; and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR⁷ optionally is substituted at one or more positions with one or moreof alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl(e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl.

In some embodiments, the disclosed substituted heterocycles may includesubstitute triazoles having a formula V:

wherein:

-   R¹ is hydrogen, or R¹ is aryl (e.g., phenyl), alkylaryl (e.g.,    benzyl), alkyldiaryl (e.g., methyldiphenyl), heteroaryl (e.g.,    N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R¹ is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), aryl (e.g., phenyl), hydroxyl, halo, cyano,    carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl;-   Alk¹ is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   n is 0 or 1;-   X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸);-   p is 0 or 1;-   Alk² is straight-chain or branched alkylenyl (e.g., a diradical of a    straight-chain or branched C₁-C₆ alkyl group);-   q is 0 or 1;-   m is 0 or 1;-   R² is hydrogen or halo, or R² is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g. benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R² is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl;-   R⁵ is hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆    alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, or halo;-   r is 0 or 1;-   R⁷ is hydrogen or halo, or R⁷ is alkyl, aryl (e.g., phenyl),    alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,    pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl,    pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,    1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,    cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl),    optionally R⁷ is substituted at one or more positions with one or    more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),    haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g.,    trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido, hydrazonyl,    carbonyl, carboxyl, and alkoxycarbonyl; and-   R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionally substituted    with halo.

In some embodiments of these disclosed substituted triazoles, at leastone of R² and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR² and R⁷ optionally are substituted at one or more positions with oneor more of alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy),haloalkyl (e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy),hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl. In some embodiments of the disclosed compounds, m is 0and R² is hydrogen, or R⁷ is hydrogen.

In some embodiments of these disclosed substituted triazoles, R² is aryl(e.g., phenyl), alkylaryl (e.g., benzyl), heteroaryl (e.g., N-pyridinyl,pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrazol-1-yl, pyrazol-3-yl,pyrazol-4-yl, 1,3-benzodioxol-5-yl, 1,3-benzodioxol-6-yl, furan-2-yl,furan-3-yl), cycloalkyl (e.g., cyclohexyl), or cycloheteroalkyl (e.g.,piperidinyl, morpholinyl), and R² optionally is substituted at one ormore positions with one or more of alkyl (e.g., C₁-C₆ alkyl), alkoxy(e.g., C₁-C₆ alkoxy), haloalkyl (e.g., trifluoromethyl), haloalkoxy(e.g., trifluoromethoxy), hydroxyl, halo, cyano, carboxyamido,hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl; and R⁷ is hydrogen.

In some embodiments of these disclosed substituted triazoles, m is 0 andR² is hydrogen; and R⁷ is aryl (e.g., phenyl), alkylaryl (e.g., benzyl),heteroaryl (e.g., N-pyridinyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl,pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, 1,3-benzodioxol-5-yl,1,3-benzodioxol-6-yl, furan-2-yl, furan-3-yl), cycloalkyl (e.g.,cyclohexyl), or cycloheteroalkyl (e.g., piperidinyl, morpholinyl), andR⁷ optionally is substituted at one or more positions with one or moreof alkyl (e.g., C₁-C₆ alkyl), alkoxy (e.g., C₁-C₆ alkoxy), haloalkyl(e.g., trifluoromethyl), haloalkoxy (e.g., trifluoromethoxy), hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl.

The formulae of the compounds disclosed herein should be interpreted asencompassing all possible stereoisomers, enantiomers, or epimers of thecompounds unless the formulae indicates a specific stereoisomer,enantiomer, or epimer. The formulae of the compounds disclosed hereinshould be interpreted as encompassing salts, esters, amides, or solvatesthereof of the compounds.

Use of the Disclosed Compounds for Inhibiting C-MYC Activity

The disclosed compounds may exhibit one or more biological activities.The disclosed compounds may inhibit binding of the MYC/Max complex toDNA (e.g., in a DNA gel shifting assay). In some embodiments, thedisclosed compounds inhibit binding of the MYC/Max complex to DNA by atleast 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% at aconcentration of less than about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM,0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM, or less. The disclosed compoundsmay not produce significant DNA damage (e.g., in an rH2AX staining assayat a concentration greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.1μM, 1.0 μM, 10 μM, 100 μM, or higher). The disclosed compounds mayinhibit the growth of cells that express c-MYC (preferably by at least50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% at a concentrationof less than about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM,0.005 μM, 0.001 μM, or less). The disclosed compounds may not inhibitthe growth of cells that do not express c-MYC (preferably by not morethan 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2% or less at a concentrationof greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0μM, 10 μM, and 100 μM or higher). Concentration ranges also arecontemplated herein, for example, a concentration range bounded byend-point concentrations selected from 0.001 μM, 0.005 μM, 0.01 μM, 0.5μM, 0.1 μM, 1.0 μM, 10 μM, and 100 μM.

The disclosed compounds may be effective in inhibiting cellproliferation of cancer cells, including cancer cells that express c-MYCand whose proliferation is inhibiting by inhibiting the biologicalactivity of c-MYC. The disclosed compounds may be effective ininhibiting cell proliferation of one or more types of cancer cellsincluding: multiple myeloma cells, such as MM.1S cells; leukemia cells,such as CCRF-CEM, HL-60(TB), MOLT-4, RPMI-8226 and SR; non-small lungcancer cells, such as A549/ATCC, EKVX, HOP-62, HOP-92, NCI-H226,NCI-H23, NCI-H322M, NCI-H460 and NCI-H522; colon cancer cells, such asCOLO 205, HCC-2998, HCT-116, HCT-15, HT29, KM12 and SW-620; CNS: SF-268,SF-295, SF-539, SNB-19, SNB-75 and U251; melanoma cancer cells, such asLOX IMVI, MALME-3M, M14, MDA-MB-435, SK-MEL-2, SK-MEL-28, SK-MEL-5,UACC-257 and UACC-62; ovarian cancer cells, such as IGR-OV1, OVCAR-3,OVCAR-4, OVCAR-5, OVCAR-8, NCI/ADR-RES and SK-OV-3; renal cancer cells,such as 786-0, A498, ACHN, CAKI-1, RXF 393, SN12C, TK-10 and UO-31;prostate cancer cells, such as DU-145 and PC-3; and breast cancer cells,such as MCF7, MDA-MB-231/ATCC, MDA-MB-468, HS 578T, BT-549 and T-47D.

Cell proliferation and inhibition thereof by the presently disclosedcompounds may be assessed by cell viability methods disclosed in the artincluding colorimetric assays that utilize dyes such as MTT, XTT, andMTS to assess cell viability. Preferably, the disclosed compounds havean IC₅₀ of less than about 10 μM, 5 μM, 1 μM, 0.5 μM, 0.01 μM, 0.005 μM,0.001 μM or lower in the selected assay.

The disclosed compounds may be formulated as anti-cancer therapeutics,including hematologic malignancies, breast, lung, pancreas and prostatemalignancies. The disclosed compounds also may be formulated asanti-inflammation therapeutics.

The compounds utilized in the methods disclosed herein may be formulatedas pharmaceutical compositions that include: (a) a therapeuticallyeffective amount of one or more compounds as disclosed herein; and (b)one or more pharmaceutically acceptable carriers, excipients, ordiluents. The pharmaceutical composition may include the compound in arange of about 0.1 to 2000 mg (preferably about 0.5 to 500 mg, and morepreferably about 1 to 100 mg). The pharmaceutical composition may beadministered to provide the compound at a daily dose of about 0.1 toabout 1000 mg/kg body weight (preferably about 0.5 to about 500 mg/kgbody weight, more preferably about 50 to about 100 mg/kg body weight).In some embodiments, after the pharmaceutical composition isadministered to a subject (e.g., after about 1, 2, 3, 4, 5, or 6 hourspost-administration), the concentration of the compound at the site ofaction may be within a concentration range bounded by end-pointsselected from 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10μM, and 100 μM (e.g., 0.1 μM-1.0 μM).

The disclosed compounds and pharmaceutical compositions comprising thedisclosed compounds may be administered in methods of treating a subjectin need thereof. For example, in the methods of treatment a subject inneed thereof may include a subject having a cell proliferative disease,disorder, or condition such as cancer (e.g., cancers such as multiplemyeloma, leukemia, non-small cell lung cancer, colon cancer, cancer ofthe central nervous system, melanoma, ovarian cancer, renal cancer,prostate cancer, and breast cancer).

In some embodiments of the disclosed treatment methods, the subject maybe administered a dose of a compound as low as 1.25 mg, 2.5 mg, 5 mg,7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg,30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg,52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg,75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 100 mg, 200 mg,500 mg, 1000 mg, or 2000 mg once daily, twice daily, three times daily,four times daily, once weekly, twice weekly, or three times per week inorder to treat the disease or disorder in the subject. In someembodiments, the subject may be administered a dose of a compound ashigh as 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg,20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg,42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg,65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg,87.5 mg, 90 mg, 100 mg, 200 mg, 500 mg, 1000 mg, or 2000 mg, once daily,twice daily, three times daily, four times daily, once weekly, twiceweekly, or three times per week in order to treat the disease ordisorder in the subject. Minimal and/or maximal doses of the compoundsmay include doses falling within dose ranges having as end-points any ofthese disclosed doses (e.g., 2.5 mg-200 mg).

In some embodiments, a minimal dose level of a compound for achievingtherapy in the disclosed methods of treatment may be at least about 10,20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450,500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400,1600, 1800, 1900, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000,15000, or 20000 ng/kg body weight of the subject. In some embodiments, amaximal dose level of a compound for achieving therapy in the disclosedmethods of treatment may not exceed about 10, 20, 30, 40, 50, 60, 70,80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700,750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 1900, 2000, 3000,4000, 5000, 6000, 7000, 8000, 9000, 10000, 15000, or 20000 ng/kg bodyweight of the subject. Minimal and/or maximal dose levels of thecompounds for achieving therapy in the disclosed methods of treatmentmay include dose levels falling within ranges having as end-points anyof these disclosed dose levels (e.g., 500-2000 ng/kg body weight of thesubject).

The compounds utilized in the methods disclosed herein may be formulatedas a pharmaceutical composition in solid dosage form, although anypharmaceutically acceptable dosage form can be utilized. Exemplary soliddosage forms include, but are not limited to, tablets, capsules,sachets, lozenges, powders, pills, or granules, and the solid dosageform can be, for example, a fast melt dosage form, controlled releasedosage form, lyophilized dosage form, delayed release dosage form,extended release dosage form, pulsatile release dosage form, mixedimmediate release and controlled release dosage form, or a combinationthereof.

The compounds utilized in the methods disclosed herein may be formulatedas a pharmaceutical composition that includes a carrier. For example,the carrier may be selected from the group consisting of proteins,carbohydrates, sugar, talc, magnesium stearate, cellulose, calciumcarbonate, and starch-gelatin paste.

The compounds utilized in the methods disclosed herein may be formulatedas a pharmaceutical composition that includes one or more bindingagents, filling agents, lubricating agents, suspending agents,sweeteners, flavoring agents, preservatives, buffers, wetting agents,disintegrants, and effervescent agents. Filling agents may includelactose monohydrate, lactose anhydrous, and various starches; examplesof binding agents are various celluloses and cross-linkedpolyvinylpyrrolidone, microcrystalline cellulose, such as Avicel® PH101and Avicel® PH102, microcrystalline cellulose, and silicifiedmicrocrystalline cellulose (ProSolv SMCC™). Suitable lubricants,including agents that act on the flowability of the powder to becompressed, may include colloidal silicon dioxide, such as Aerosil®200,talc, stearic acid, magnesium stearate, calcium stearate, and silicagel. Examples of sweeteners may include any natural or artificialsweetener, such as sucrose, xylitol, sodium saccharin, cyclamate,aspartame, and acsulfame. Examples of flavoring agents are Magnasweet®(trademark of MAFCO), bubble gum flavor, and fruit flavors, and thelike. Examples of preservatives may include potassium sorbate,methylparaben, propylparaben, benzoic acid and its salts, other estersof parahydroxybenzoic acid such as butylparaben, alcohols such as ethylor benzyl alcohol, phenolic compounds such as phenol, or quaternarycompounds such as benzalkonium chloride.

Suitable diluents may include pharmaceutically acceptable inert fillers,such as microcrystalline cellulose, lactose, dibasic calcium phosphate,saccharides, and mixtures of any of the foregoing. Examples of diluentsinclude microcrystalline cellulose, such as Avicel® PH101 and Avicel®PH102; lactose such as lactose monohydrate, lactose anhydrous, andPharmatose® DCL21; dibasic calcium phosphate such as Emcompress®;mannitol; starch; sorbitol; sucrose; and glucose.

Suitable disintegrants include lightly crosslinked polyvinylpyrrolidone, corn starch, potato starch, maize starch, and modifiedstarches, croscarmellose sodium, cross-povidone, sodium starchglycolate, and mixtures thereof.

Examples of effervescent agents are effervescent couples such as anorganic acid and a carbonate or bicarbonate. Suitable organic acidsinclude, for example, citric, tartaric, malic, fumaric, adipic,succinic, and alginic acids and anhydrides and acid salts. Suitablecarbonates and bicarbonates include, for example, sodium carbonate,sodium bicarbonate, potassium carbonate, potassium bicarbonate,magnesium carbonate, sodium glycine carbonate, L-lysine carbonate, andarginine carbonate. Alternatively, only the sodium bicarbonate componentof the effervescent couple may be present.

The compounds utilized in the methods disclosed herein may be formulatedas a pharmaceutical composition for delivery via any suitable route. Forexample, the pharmaceutical composition may be administered via oral,intravenous, intramuscular, subcutaneous, topical, and pulmonary route.Examples of pharmaceutical compositions for oral administration includecapsules, syrups, concentrates, powders and granules. In someembodiments, the compounds are formulated as a composition foradministration orally (e.g., in a solvent such as 5% DMSO in oil such asvegetable oil).

The compounds utilized in the methods disclosed herein may beadministered in conventional dosage forms prepared by combining theactive ingredient with standard pharmaceutical carriers or diluentsaccording to conventional procedures well known in the art. Theseprocedures may involve mixing, granulating and compressing or dissolvingthe ingredients as appropriate to the desired preparation.

Pharmaceutical compositions comprising the compounds may be adapted foradministration by any appropriate route, for example by the oral(including buccal or sublingual), rectal, nasal, topical (includingbuccal, sublingual or transdermal), vaginal or parenteral (includingsubcutaneous, intramuscular, intravenous or intradermal) route. Suchformulations may be prepared by any method known in the art of pharmacy,for example by bringing into association the active ingredient with thecarrier(s) or excipient(s).

Pharmaceutical compositions adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilliquid emulsions.

Pharmaceutical compositions adapted for transdermal administration maybe presented as discrete patches intended to remain in intimate contactwith the epidermis of the recipient for a prolonged period of time. Forexample, the active ingredient may be delivered from the patch byiontophoresis.

Pharmaceutical compositions adapted for topical administration may beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, impregnated dressings, sprays, aerosols or oilsand may contain appropriate conventional additives such aspreservatives, solvents to assist drug penetration and emollients inointments and creams.

For applications to the eye or other external tissues, for example themouth and skin, the pharmaceutical compositions are preferably appliedas a topical ointment or cream. When formulated in an ointment, thecompound may be employed with either a paraffinic or a water-miscibleointment base. Alternatively, the compound may be formulated in a creamwith an oil-in-water cream base or a water-in-oil base. Pharmaceuticalcompositions adapted for topical administration to the eye include eyedrops where the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent.

Pharmaceutical compositions adapted for nasal administration where thecarrier is a solid include a coarse powder having a particle size (e.g.,in the range 20 to 500 microns) which is administered in the manner inwhich snuff is taken (i.e., by rapid inhalation through the nasalpassage from a container of the powder held close up to the nose).Suitable formulations where the carrier is a liquid, for administrationas a nasal spray or as nasal drops, include aqueous or oil solutions ofthe active ingredient.

Pharmaceutical compositions adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe formulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets.

Tablets and capsules for oral administration may be in unit dosepresentation form, and may contain conventional excipients such asbinding agents, for example syrup, acacia, gelatin, sorbitol,tragacanth, or polyvinylpyrrolidone; fillers, for example lactose,sugar, maize-starch, calcium phosphate, sorbitol or glycine; tablettinglubricants, for example magnesium stearate, talc, polyethylene glycol orsilica; disintegrants, for example potato starch; or acceptable wettingagents such as sodium lauryl sulphate. The tablets may be coatedaccording to methods well known in normal pharmaceutical practice. Oralliquid preparations may be in the form of, for example, aqueous or oilysuspensions, solutions, emulsions, syrups or elixirs, or may bepresented as a dry product for reconstitution with water or othersuitable vehicle before use. Such liquid preparations may containconventional additives, such as suspending agents, for example sorbitol,methyl cellulose, glucose syrup, gelatin, hydroxyethyl cellulose,carboxymethyl cellulose, aluminium stearate gel or hydrogenated ediblefats, emulsifying agents, for example lecithin, sorbitan monooleate, oracacia; non-aqueous vehicles (which may include edible oils), forexample almond oil, oily esters such as glycerine, propylene glycol, orethyl alcohol; preservatives, for example methyl or propylp-hydroxybenzoate or sorbic acid, and, if desired, conventionalflavoring or coloring agents.

Combination Therapies and Pharmaceutical Compositions

The disclosed compounds or pharmaceutical compositions comprising thedisclosed compounds may be administered in methods of treatment. Forexample, the disclosed compounds or pharmaceutical compositionscomprising the disclosed compounds may be administered in methods oftreating cell proliferative diseases and disorders. Cell proliferativediseases and disorders treated by the disclosed methods may include, butare not limited to, cancers selected from the group consisting ofmultiple myeloma, leukemia, non-small cell lung cancer, colon cancer,cancer of the central nervous system, melanoma, ovarian cancer, renalcancer, prostate cancer, and breast cancer.

Optionally, the disclosed compounds or pharmaceutical compositionscomprising the disclosed compounds may be administered with additionaltherapeutic agents, optionally in combination, in order to treat cellproliferative diseases and disorders. In some embodiments of thedisclosed methods, one or more additional therapeutic agents areadministered with the disclosed compounds or with pharmaceuticalcompositions comprising the disclosed compounds, where the additionaltherapeutic agent is administered prior to, concurrently with, or afteradministering the disclosed compounds or the pharmaceutical compositionscomprising the disclosed compounds. In some embodiments, the disclosedpharmaceutical composition are formulated to comprise the disclosedcompounds and further to comprise one or more additional therapeuticagents, for example, one or more additional therapeutic agents fortreating cell proliferative diseases and disorders.

EXAMPLES

The following Examples are illustrative and are not intended to limitthe scope of the claimed subject matter.

Example 1—Identification of Small Molecule Inhibitors of C-MYC DNABinding Activity

Introduction

MYC is the most frequently amplified oncogene in human cancers. It hasbeen extensively validated as essential for tumor initiation andmaintenance in numerous tumor histologies. Numerous studies haveprovided solid evidence that pharmacologic targeting of MYC woulddirectly affect tumor progression. One example is OmoMYC, adominant-negative peptide of MYC that competitively binds MYC in amanner that prevents MYC-Max heterodimerization. OmoMYC expressionprompts rapid growth arrest and down-regulation of MYC target genes incancer cells both in vitro and in vivo. Small molecule inhibitors of MYCwill be the optimal form for drug development. However, disruption ofMYC-Max interactions through small molecules has been difficult becausethere are no obvious binding regions in the interface. Thus far, over 30small molecules have been documented with MYC inhibition activity invitro, but the evidence for their in vivo activities is lacking, likelydue to their poor drug-like properties. Among these compounds, 10058-f4and 10075-G5 are well-known for their specificities and relatively clearmechanisms in interrupting MYC-Max binding. However, the in vivo studieswere quite disappointing because of their rapid metabolism. Thus,developing new MYC inhibitors with high potency and specificity as wellas favorable drug-like properties will be critical to effectively targetMYC.

To this end, we carried out an in silico screen to identify compoundsthat might inhibit the binding of c-MYC to DNA. These compounds weretested in several cell-based assays to identify the most active hits.The best hit, Min-9 (NUCC-176234) and its related analogs were shown toprevent c-MYC/DNA binding. We then synthesized a series of novelstructural analogs and these were tested in the same c-MYC-relevantassays. Our new compounds display excellent potency at inhibitingc-MYC/DNA binding. The compounds we have developed using a novelapproach possess greatly improved drug-like properties over existingsmall molecules such as 10058-f4 and therefore represent excellentstarting points for developing MYC-targeting therapeutics.

Results

In the absence of a regular small-molecule ligand-binding pocket in thec-MYC/Max/DNA ternary complex, we applied multiple independent in silicoapproaches to increase our likelihood of successfully identifying newsmall molecule inhibitors. (See FIG. 1). We carried out in-silicoscreening of a 10 million compound drug-like library. We applied twodifferent approaches to screen the ZINC compound database after removingpromiscuous and non drug-like compounds using PAINS filters. The firstapproach is based on a 3-tier docking protocol using a published crystalstructure of MYC/Max bound to DNA. After defining a putativeligand-binding site as reported in the literature, the compound librarywas screened using the docking tool. The second approach was based onbuilding a pharmacophore model considering of 32 compounds reported toinhibit MYC and screening the Zinc database against this pharmacophore.We obtained 69 hits from the structure-based screen and 60 hits from theligand-based pharmacophore screen, with 32 compounds in common betweenthe two approaches.

To test the compounds, we evaluated the in silico hits in a MYC E-Boxluciferase reporter assay to measure the effects of these compounds(referred to as Min-1 to Min-32) on MYC transcriptional activity. Asshown in FIG. 2, about 10 compounds have similar or better activitycompared to positive control 10058-F4 at 25 μM. (See FIG. 2).

We next examined the ability of the compounds to selectively inhibit theproliferation of wild type cells expressing MYC relative to cells withMYC knockout. We tested the top 13 active compounds in the first screenassay. FIG. 3 shows a graph of growth inhibition by each compound on thewild type and MYC knockout rat fibroblasts at the dose with the greatestselectivity. More than half of the tested compounds show better growthinhibitory effect on MYC WT compared to MYC KO cells. Min9-S7(NUCC-0176248) is very promising because of its low effectiveconcentration (6 μM) and high specificity. Min9-S9 (NUCC-0176250) alsoshows a great selectivity at an acceptable dosage (50 μM).

Min9 (NUCC-0176234) was also tested in a cell viability assay against acMYC wild-type (WT) and a cMYC KO line. As shown in FIG. 4, thiscompound reduces cell viability much more in the WT line than the KOcells, indicating a mechanism directly related to cMYC.

We also tested our best hit compound Min9 (NUCC-0176234) and newlysynthesized analogs for effects of these compounds on MYC/Max binding toDNA in electrophoretic mobility shift assays (EMSAs). (See FIG. 5a andFIG. 5b ). We expected the active compounds to impair MYC/Max binding toDNA. Several structural analogs of Min9 were tested over multiple dosesfor inhibiting MYC-DNA binding and we observed a dose-dependentinhibition. (See FIG. 5c ).

Min9 (NUCC-0176234) was also tested for its ability to cause DNA damagein an rH2AX staining assay. We would not expect cMYC-targeting agents toproduce significant DNA damage. Compounds that act directly against DNAsuch as doxorubicin do however. We observed essentially no DNA damagecaused by Min9 (NUCC-0176234). (See FIG. 6).

In vitro metabolism of NUCC-176242 and NUCC-176248 were tested usingmouse liver microsomes and a mouse S9 fraction. (See FIG. 9).NUCC-176242 was significantly metabolism by the mouse S9 fraction versusNUCC-176248 likely due to S9 conjugation at the N-1 nitrogen atom of thepyrazole ring.

The pharmacokinetics of NUCC-176242 and NUCC-176248 were studied in miceby administering a dose of 5 mg/kg intravenously and measuring theplasma concentration versus time. (See FIG. 10). The observed in vivometabolism of NUCC-176242 and NUCC-176248 correlated well with theobserved in vitro metabolism tested above for of NUCC-176242 andNUCC-176248.

Chemistry

General Experimental.

All chemical reagents were obtained from commercial suppliers and usedwithout further purification unless otherwise stated. Anhydrous solventswere purchased from Sigma-Aldrich, and dried over 3 Å molecular sieveswhen necessary. DCM and THE were purified by passage through a bed ofactivated alumina. Normal-phase flash column chromatography wasperformed using Biotage KP-Sil 50 μm silica gel columns and ACS gradesolvents on a Biotage Isolera flash purification system. Analytical thinlayer chromatography (TLC) was performed on EM Reagent 0.25 mm silicagel 60 F₂₅₄ plates and visualized by UV light or iodine vapor. Liquidchromatography/mass spectrometry (LCMS) was performed on a WatersAcquity-H UPLC system with a 2.1 mm×50 mm, 1.7 μm, reversed phase BEHC18 column and LCMS grade solvents. A gradient elution from 95%water+0.1% formic acid/5% acetonitrile+0.1% formic acid to 95%acetonitrile+0.1% formic acid/5% water+0.1% formic acid over 2 min plusa further minute continuing this mixture at a flow rate of 0.85 mL/minwas used as the eluent. Total ion current traces were obtained forelectrospray positive and negative ionization (ESI+/ESI−). Proton (H)and carbon (¹³C) NMR spectra were recorded on a Bruker Avance IIIw/direct cryoprobe spectrometer. Chemical shifts were reported in ppm(δ) and were referenced using residual non-deuterated solvent as aninternal standard. The chemical shifts for ¹H NMR and ¹³C NMR arereported to the second decimal place. Proton coupling constants areexpressed in hertz (Hz). The following abbreviations were used to denotespin multiplicity for proton NMR: s=singlet, d=doublet, t=triplet,q=quartet, m=multiplet, brs=broad singlet, dd=doublet of doublets,dt=doublet of triplets, quin=quintet, tt=triplet of triplets. In somecases, overlapping signals occurred in the ¹³C NMR spectra.

Representative Examples for General Synthetic Method

A. Synthetic Method A

Synthetic Method A

Step 1.

Over a solution of the phenol (1 equiv.), in 100 mL of acetone,bromoacetonitrile (1 equiv.) were added followed by potassium carbonate(1.5 equiv.) were added. Then, the solution stirred at 60° C. for 4.5 h.The reaction was quenched by adding 30 mL of NaHCO₃ aqueous solution andwater, extracted with of EtOAc (3×30 mL). The combined organic layerswere washed with brine and dried over anhydrous Na₂SO₄. Solvent was thenremoved under reduced pressure and the residue was purified by silicagel chromatography.

Step 2.

Over a cold ice-bath solution of the acetonitrile-phenoxy intermediate(1 equiv.) in 6.5 mL of benzene (1M solution), a HCL solution in dioxane(10 equiv.) was added dropwise. The resulting solution stirred for 1 hbefore a resorcinol (1 equiv.) and ZnCl₂ (1 equiv.) in 10 mL of diethylether was added slowly. The resulting solution stirred from 0° C. to RTfor 16 h. The resulting suspension was centrifuged and the solid wasseparated. The solid was washed with water and dried under vacuum.

Step 3.

A suspension of the O-phenoxy-acetophenone (1 mmol) with TFFA (5 equiv)and pyridine (5 equiv.) was heated at 120° C. for 4 h. Then, it was cooldown to r.t. In some cases, product was precipitated, in others waterwas added and then extracted with EtOAc (3×10 mL). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtrated andconcentrated. The residue was purified by silica gel chromatography.

Step 4.

A suspension of the phenoxy-chromenone, (1 equiv.), K₂CO₃ (2 equiv.) andthe halo alkane (1.1 equiv.) in 5 mL of acetone (0.8 M) was heated at60° C. for 16 h. The reaction was filtered through a funnel and thesolvent removed under reduced pressure. The crude residue was trituratedwith water and dried under reduced pressure until dryness.

Step 5.

A solution of the previous phenoxy-chromenone (1 equiv.) with thedesired hydrazine (3 equiv.) in 2 mL of EtOH (0.1M) was heated at 70° C.for 45 minutes. The solution was cooled down to room temperature andconcentrated. The solid residue was directly purified by silica gelchromatography (n-hexanes/ethyl acetate=5:1 to 1:1) providing thedesired pyrazole.

Examples

NUCC-200683: ¹H NMR (500 MHz, CDCl₃) δ 7.81 (dd, J=7.9, 1.6 Hz, 1H),7.18 (td, J=7.7, 1.6 Hz, 1H), 7.10-7.02 (m, 2H), 6.92-6.86 (m, 2H),6.86-6.79 (m, 2H), 2.56 (q, J=7.6 Hz, 2H), 1.17 (t, J=7.6 Hz, 3H) ppm.

NUCC-198411: ¹H NMR (500 MHz, CDCl₃) δ 7.00-6.92 (m, 3H), 6.80-6.68 (m,2H), 6.44-6.38 (m, 1H), 6.32 (d, J=8.4 Hz, 1H), 2.52 (q, J=7.8 Hz, 2H),1.13 (t, J=7.4 Hz, 3H) ppm.

NUCC-198406: ¹H NMR (500 MHz, CDCl₃) δ 7.64 (d, J=8.6 Hz, 1H), 7.06 (d,J=8.6 Hz, 2H), 6.82 (d, J=8.6 Hz, 2H), 6.41 (d, J=2.4 Hz, 1H), 6.33 (dd,J=8.6, 2.4 Hz, 1H), 2.56 (q, J=7.5 Hz, 2H), 1.17 (t, J=7.7 Hz, 3H) ppm.

NUCC-196355: ¹H NMR (500 MHz, CDCl₃) δ 7.69 (d, J=8.7 Hz, 1H), 7.47 (d,J=2.0 Hz, 1H), 7.42 (d, J=8.3 Hz, 1H), 7.19 (dd, J=8.3, 2.0 Hz, 1H),7.07 (d, J=8.7 Hz, 2H), 6.86-6.76 (m, 2H), 6.48 (d, J=13.3 Hz, 2H), 4.95(s, 2H), 2.57 (q, J=7.6 Hz, 2H), 1.18 (t, J=7.6 Hz, 3H) ppm.

NUCC-196342: ¹H NMR (500 MHz, CDCl₃) δ 7.69 (d, J=8.8 Hz, 1H), 7.43 (dd,J=7.7, 1.5 Hz, 1H), 7.35-7.27 (m, 4H), 7.06 (ddd, J=8.3, 7.7, 1.6 Hz,1H), 6.96 (td, J=7.6, 1.4 Hz, 1H), 6.65 (dd, J=8.3, 1.5 Hz, 1H), 6.50(d, J=14.5 Hz, 2H), 4.97 (s, 2H) ppm.

NUCC-196295: ¹H NMR (400 MHz, CDCl₃) δ 7.70 (d, J=8.7 Hz, 1H), 7.37 (s,1H), 7.28 (dd, J=5.4, 1.2 Hz, 2H), 7.07 (d, J=8.7 Hz, 2H), 6.86-6.79 (m,2H), 6.50 (s, 2H), 4.98 (s, 2H), 2.57 (q, J=7.6 Hz, 1H), 1.18 (t, J=7.6Hz, 2H) ppm.

B. Synthetic Method B

Synthetic Method B

Step 1.

Over a suspension of 1-(2,4-dihydroxyphenyl)ethan-1-one (5.00 g, 32.89mmol, 1 equiv.) in trifluoroacetic anhydride (18.50 mL, 131.56 mmol, 4equiv.) placed in a high-pressure tube, sodium 2,2,2-trifluoroacetate(9.84 g, 72.36 mmol, 2.2 equiv.) was added and the system was capped andstirred at 110° C. for 24 h. The reaction was allowed to cool down toapproximately 70° C. and then was diluted with 200 mL of EtOAc. Themixture was neutralized by adding saturated aqueous K₂CO₃ solution untilno more bubbling was observed. Layers were separated and the aqueousphase was extracted with more EtOAc (3×150 mL). The combined organiclayers were washed with brine and dried over anhydrous Na₂SO₄. Thesolution was then concentrated to 100-150 mL of EtOAc. Then the flaskwas capped and kept at room temperature for 1-2 days, obtaining a solidwhich was filtrated and dried under vacuum to obtain 4.09 g of pure 1 asa white solid in 54% yield.

Then, over a solution of the solid obtained (4 g, 17.31 mmol, 1 equiv.)and iodine (17.57 g, 69.24 mmol, 4 equiv.) in 110 mL of CHCl₃, pyridine(5.59 mL, 69.24 mmol, 4 equiv.) were added. The resulting solution wasstirred at room temperature for 16 h. Then, 120 mL of saturated aqueousNa₂S₂O₃ were added and the resulting mixture stirred for one hour. Theorganic layer was separated and the aqueous phase was extracted withCH₂Cl₂ (3×100 mL). The combined organic layers were washed with brineand dried over anhydrous Na₂SO₄. Solvent was then removed under reducedpressure and the residue was triturated with diethyl ether several timesto obtain a pale white solid in 90% yield (5.55 g, 15.60 mmol): mp205-206° C. ¹H NMR (500 MHz, CDCl₃) δ 8.11 (dd, J=8.9, 1.0 Hz, 1H), 7.15(dd, J=8.9, 1.0 Hz, 1H), 6.77-6.69 (m, 1H), 6.36 (s, 1H) ppm. ¹³C NMR(126 MHz, CDCl₃) δ 175.8, 161.5, 156.0, 152.5 (q, ²J=39.1 Hz), 128.1,119.0, 118.6 (q, ¹J=272.2 Hz), 117.5, 114.7, 110.9 ppm. LRMS (EI): masscalc for C₁₀H₅F₃IO3⁺ [M+H]⁺=356.9, found=357.1.

Step 2.

A suspension of 7-hydroxy-3-iodo-2-(trifluoromethyl)-4H-chromen-4-one,(1 g, 2.8 mmol, 1 equiv.), the halo-alkane (3.4 mmol, 1.2 equiv.) andK₂CO₃ (0.77 g, 5.6 mmol, 2 equiv.) in 5 mL of acetone was heated at 60°C. for 16 h. The reaction was filtered through a funnel and the solventremoved under reduced pressure. The crude residue was triturated withwater and dried under reduced pressure until dryness.

Step 3.

A suspension of the previous alkylated chromenone (0.34 mmol, 1 equiv.),with the corresponding boronic acid (0.37 mmol, 1.1 equiv.), Na₂CO₃(0.68 mmol, 2 equiv.) and Pd(dppf)Cl₂ (0.026 mmol, 0.08 equiv.) in 3.5mL of a mixture 1:2:6 of EtOH:water:toluene was bubbled with nitrogengas for 10 minutes. Then, the flask was capped, and the mixture washeated at 90° C. for 2 h. The dark solution was cool down to roomtemperature and diluted with EtOAc. The organic layer was separated andthe aqueous phase was extracted with EtOAc (3×3 mL). The combinedorganic layers were washed with brine and dried over anhydrous Na₂SO₄.Solvent was then removed under reduced pressure and the residue waspurified by silica gel chromatography.

Step 4.

A solution of the previous chromenone (0.2 mmol, 1 equiv.) with thedesired hydrazine (0.6 mmol, 3 equiv.) in 2 mL of EtOH was heated at 70°C. for 45 minutes. The solution was cooled down to room temperature andconcentrated. The solid residue was directly purified by silica gelchromatography (n-hexanes/ethyl acetate=5:1 to 1:1) providing thedesired pyrazole.

Examples

NUCC-201634: ¹H NMR (500 MHz, CDCl₃) δ 7.75 (d, J=2.2 Hz, 1H), 7.58 (dd,J=15.0, 8.5 Hz, 2H), 7.52 (dd, J=8.5, 2.2 Hz, 1H), 6.87 (s, 1H), 6.65(d, J=8.8 Hz, 1H), 3.97 (t, J=6.5 Hz, 2H), 1.63-1.47 (m, 5H), 0.84 (d,J=6.5 Hz, 6H) ppm.

NUCC-201632: ¹H NMR (500 MHz, MeOH-d₄) δ 8.82 (d, J=2.1 Hz, 1H), 8.77(d, J=2.1 Hz, 1H), 8.13 (t, J=2.1 Hz, 1H), 7.33-7.25 (m, 2H), 7.23 (d,J=8.6 Hz, 1H), 7.20-7.14 (m, 2H), 6.77 (d, J=8.6 Hz, 1H), 6.56 (s, 1H),5.08 (s, 2H), 3.80 (s, 3H) ppm.

NUCC-201227: ¹H NMR (500 MHz, CDCl₃) δ 10.59 (s, 1H), 7.80 (d, J=2.1 Hz,1H), 7.57 (dd, J=8.2, 2.1 Hz, 1H), 7.51 (d, J=8.2 Hz, 1H), 7.47 (d,J=8.7 Hz, 1H), 6.90 (s, 1H), 6.58 (d, J=8.7 Hz, 1H), 3.97 (s, 3H), 3.94(t, J=6.3 Hz, 2H), 1.61 (dq, J=7.9, 6.4 Hz, 2H), 1.33 (h, J=7.5 Hz, 2H),0.87 (t, J=7.4 Hz, 3H) ppm.

NUCC-201226: ¹H NMR (500 MHz, CDCl₃) δ 7.75 (d, J=2.1 Hz, 1H), 7.59 (d,J=8.3 Hz, 1H), 7.50 (dd, J=8.3, 2.1 Hz, 1H), 7.18 (d, J=8.5 Hz, 1H),6.66 (d, J=8.5 Hz, 1H), 6.54 (s, 1H), 3.96 (t, J=6.3 Hz, 2H), 3.82 (s,3H), 1.68-1.58 (m, 2H), 1.36-1.29 (m, 2H), 0.87 (t, J=7.4 Hz, 3H) ppm.

NUCC-0201213: ¹H NMR (500 MHz, CDCl₃) δ 8.81 (dd, J=20.5, 2.0 Hz, 2H),8.03 (d, J=1.9 Hz, 1H), 7.29-7.15 (m, 3H), 6.70 (d, J=8.6 Hz, 1H), 6.59(s, 1H), 5.50 (s, 1H), 4.13-3.71 (m, 5H), 1.78-1.31 (m, 5H), 0.87 (d,J=6.3 Hz, 7H) ppm.

NUCC-201208: ¹H NMR (500 MHz, CDCl₃) δ 8.83 (d, J=2.1 Hz, 1H), 8.79 (d,J=2.1 Hz, 1H), 8.04 (t, J=2.1 Hz, 1H), 7.21 (d, J=8.7 Hz, 1H), 6.70 (d,J=8.7 Hz, 1H), 6.59 (s, 1H), 4.01 (t, J=6.4 Hz, 2H), 3.83 (s, 3H),1.71-1.43 (m, 3H), 0.87 (d, J=6.4 Hz, 6H) ppm.

NUCC-0201207: ¹H NMR (500 MHz, CDCl₃) δ 8.93-8.66 (m, 2H), 8.04 (d,J=2.1 Hz, 1H), 7.34-7.12 (m, 2H), 6.82-6.45 (m, 3H), 5.85 (s, 1H),4.10-3.71 (m, 7H), 1.65 (q, J=6.8 Hz, 3H), 1.40-1.01 (m, 9H), 1.01-0.58(m, 4H) ppm.

NUCC-201206: ¹H NMR (500 MHz, CDCl₃) δ 8.82 (s, 2H), 8.01 (d, J=2.3 Hz,1H), 7.21 (d, J=8.6 Hz, 1H), 6.70 (d, J=8.6 Hz, 1H), 6.58 (s, 1H), 4.00(t, J=6.3 Hz, 2H), 3.84 (s, 3H), 1.65-1.53 (m, 3H), 0.84 (dd, J=6.5, 2.6Hz, 6H) ppm.

NUCC-0201205: ¹H NMR (500 MHz, CDCl₃) δ 8.74 (d, J=43.1 Hz, 2H), 8.04(s, 1H), 7.30-7.14 (m, 2H), 6.74-6.48 (m, 2H), 3.96 (t, J=6.1 Hz, 3H),3.83 (d, J=3.6 Hz, 2H), 1.62 (q, J=6.7 Hz, 3H), 1.38-1.06 (m, 8H), 0.82(t, J=6.8 Hz, 4H) ppm.

NUCC-0201204: ¹H NMR (500 MHz, CDCl₃) δ 8.84-8.61 (m, 2H), 8.04 (s, 1H),7.31-7.14 (m, 1H), 6.67 (d, J=8.6 Hz, 1H), 6.56 (s, 1H), 3.97 (t, J=6.2Hz, 2H), 3.83 (s, 2H), 1.69-1.54 (m, 2H), 1.30 (q, J=7.5 Hz, 2H), 0.85(t, J=7.5 Hz, 3H) ppm.

NUCC-0201201: 1H NMR (500 MHz, CDCl₃) δ 8.05-7.74 (m, 5H), 7.57 (d,J=8.8 Hz, 1H), 7.09-6.85 (m, 4H), 6.75 (d, J=8.7 Hz, 1H), 6.59 (d, J=8.1Hz, 3H), 4.95 (s, 2H), 2.90 (d, J=37.0 Hz, 2H) ppm.

NUCC-0201198: ¹H NMR (500 MHz, CDCl₃) δ 10.79 (s, 1H), 7.89 (d, J=40.7Hz, 3H), 7.57 (d, J=8.7 Hz, 1H), 6.92 (s, 1H), 6.64 (d, J=8.8 Hz, 1H),3.95 (t, J=6.2 Hz, 2H), 1.77-1.49 (m, 2H), 1.46-1.06 (m, 7H), 0.81 (t,J=6.8 Hz, 3H) ppm.

NUCC-0201197: ¹H NMR (500 MHz, CDCl₃) δ 8.08-7.75 (m, 1H), 3.92 (t,J=6.3 Hz, 1H), 1.65 (q, J=6.8 Hz, 1H), 0.87 (t, J=7.4 Hz, 1H) ppm.

NUCC-0201196: ¹H NMR (500 MHz, Chloroform-d) δ 10.76 (s, 1H), 7.88 (d,J=35.4 Hz, 3H), 7.57 (d, J=8.7 Hz, 1H), 6.91 (s, 1H), 6.65 (d, J=8.7 Hz,1H), 3.98 (t, J=6.3 Hz, 2H), 1.54 (dq, J=37.9, 6.7 Hz, 3H), 0.83 (d,J=6.5 Hz, 7H) ppm.

NUCC-0201195: ¹H NMR (500 MHz, CDCl₃) δ 11.63 (s, 1H), 8.96 (s, 1H),8.77 (s, 1H), 8.12 (s, 1H), 7.57 (d, J=8.7 Hz, 1H), 6.95 (s, 1H), 6.63(d, J=8.7 Hz, 1H), 4.00 (t, J=6.5 Hz, 2H), 1.59 (ddt, J=32.4, 13.3, 6.6Hz, 6H), 0.87 (d, J=6.4 Hz, 7H) ppm.

NUCC-0201193: ¹H NMR (500 MHz, CDCl₃) δ 11.25 (s, 1H), 8.93 (d, J=2.1Hz, 1H), 8.78 (d, J=2.0 Hz, 1H), 8.11 (d, J=2.1 Hz, 1H), 7.57 (d, J=8.8Hz, 1H), 6.96 (s, 1H), 6.63 (d, J=8.7 Hz, 1H), 3.98 (t, J=6.4 Hz, 2H),1.81-1.45 (m, 7H), 1.32 (p, J=7.4 Hz, 2H), 0.89 (t, J=7.4 Hz, 4H) ppm.

NUCC-0201192: ¹H NMR (500 MHz, CDCl₃) δ 11.64 (s, 2H), 8.87 (d, J=77.5Hz, 2H), 8.10 (s, 1H), 7.57 (d, J=8.7 Hz, 1H), 6.94 (s, 1H), 6.63 (d,J=8.7 Hz, 1H), 3.96 (t, J=6.3 Hz, 1H), 1.82-1.40 (m, 10H), 1.40-1.07 (m,7H), 0.83 (t, J=6.7 Hz, 3H) ppm.

NUCC-0201039: ¹H NMR (500 MHz, MeOH-d₄) δ 8.50 (s, 1H), 7.83 (d, J=2.6Hz, 1H), 7.67 (d, J=8.5 Hz, 1H), 7.38-7.24 (m, 2H), 7.19 (d, J=8.1 Hz,2H), 7.04-6.91 (m, 1H), 6.79 (d, J=8.7 Hz, 1H), 5.22-4.99 (m, 2H), 4.41(dd, J=11.3, 6.9 Hz, 2H), 2.05 (s, 2H), 1.26 (t, J=7.0 Hz, 3H) ppm; ¹³CNMR (126 MHz, MeOD) δ 157.2, 135.8, 133.1, 128.6, 128.1, 127.9, 125.3,101.3, 101.3, 69.1, 62.4, 13.3 ppm.

NUCC-0201037: ¹H NMR (500 MHz, CDCl₃) δ 8.70 (d, J=48.4 Hz, 2H), 8.01(s, 1H), 7.56-7.38 (m, 1H), 7.17 (td, J=5.1, 2.3 Hz, 2H), 7.04 (dd,J=8.1, 3.8 Hz, 2H), 6.80 (s, 1H), 6.67-6.55 (m, 1H), 4.94 (d, J=4.0 Hz,2H) ppm.

NUCC-0201036: ¹H NMR (500 MHz, MeOH-d₄) δ 6.49 (d, J=1.8 Hz, 1H), 6.42(d, J=7.7 Hz, 1H), 6.29-6.09 (m, 3H), 5.83-5.65 (m, 4H), 5.47 (s, 1H),5.32 (d, J=8.7 Hz, 1H), 3.52 (s, 2H), 3.35 (s, 7H), 1.81 (s, 1H), 1.54(s, 3H); ¹³C NMR (126 MHz, Methanol-d₄) δ 161.71, 155.20, 134.93,134.06, 131.66, 128.50, 126.96, 126.72, 126.63, 99.87, 67.78, 41.46 ppm.

NUCC-0201031: ¹H NMR (500 MHz, CDCl₃) δ 7.89 (d, J=5.8 Hz, 2H), 7.76 (d,J=5.6 Hz, 1H), 7.64-7.36 (m, 1H), 6.82 (d, J=5.9 Hz, 1H), 6.56 (dd,J=10.6, 4.7 Hz, 1H), 3.85 (q, J=6.1, 4.7 Hz, 2H), 3.24 (s, 2H), 1.59 (p,J=6.8 Hz, 2H), 0.99-0.65 (m, 3H); ¹³C NMR (126 MHz, CDCl₃) δ 157.1,135.8, 131.7, 127.9, 124.6, 122.5, 120.5, 109.9, 103.9, 100.9, 70.04,49.6, 49.4, 49.2, 49.1, 48.9, 48.7, 48.57, 22.3, 10.2 ppm.

NUCC-0201027: ¹H NMR (500 MHz, CDCl₃) δ 7.88 (d, J=5.1 Hz, 3H), 7.55 (d,J=8.6 Hz, 1H), 7.35-7.16 (m, 1H), 7.03-6.80 (m, 3H), 6.80-6.66 (m, 2H),6.61 (d, J=8.7 Hz, 1H), 4.11 (t, J=6.5 Hz, 2H), 3.74 (d, J=1.5 Hz, 3H),2.84 (t, J=6.6 Hz, 2H) ppm; ¹³C NMR (126 MHz, CDCl₃) δ 158.2, 157.1,131.6, 129.8, 129.7, 128.2, 124.5, 122.4, 115.6, 113.7, 109.3, 101.5,69.7, 55.2, 34.6 ppm.

NUCC-0201025: ¹H NMR (500 MHz, CDCl₃) δ 8.00 (d, J=39.3 Hz, 3H),7.43-7.29 (m, 1H), 7.29-7.14 (m, 2H), 7.04-6.93 (m, 2H), 6.86-6.75 (m,1H), 6.70 (s, 1H), 5.28 (s, 1H), 4.28 (t, J=6.6 Hz, 2H), 3.94 (s, 2H),3.01 (q, J=6.6 Hz, 2H) ppm; ¹³C NMR (126 MHz, CDCl₃) δ 157.8, 156.9,151.4, 142.1, 141.8, 139.5, 136.3, 136.2, 134.7, 132.4, 132.3, 132.04,131.7, 131.5, 131.4, 131.2, 130.1, 130.05, 128.4, 128.4, 128.1, 126.6,124.4, 122.3, 122.2, 121.5, 121.5, 121.5, 120.05, 115.7, 115.4, 109.5,109.3, 105.4, 104.7, 104.2, 101.6, 69.3, 69.1, 37.7, 34.9, 34.8 ppm.

NUCC-200736: ¹H NMR (500 MHz, CDCl₃) δ 7.82 (d, J=1.8 Hz, 1H), 7.63 (dd,J=8.0, 1.8 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 7.28-7.20 (m, 3H), 7.11 (dd,J=8.5, 3.5 Hz, 2H), 6.68 (d, J=8.5 Hz, 1H), 6.55 (s, 1H), 5.04 (d, J=2.6Hz, 2H), 3.81 (s, 3H) ppm.

NUCC-200723: ¹H NMR (500 MHz, CDCl₃) δ 13.15 (s, 1H), 7.96-7.81 (m, 1H),7.72 (d, J=8.9 Hz, 1H), 7.13 (d, J=8.5 Hz, 1H), 6.66 (d, J=8.5 Hz, 1H),6.59 (s, 1H), 6.52 (d, J=8.9 Hz, 1H), 3.84 (s, 3H) ppm.

NUCC-200576: ¹H NMR (500 MHz, MeOH-d₄) δ 7.49-7.44 (m, 2H), 7.42-7.37(m, 2H), 7.34-7.30 (m, 1H), 7.30-7.24 (m, 1H), 7.13 (d, J=8.4 Hz, 1H),7.05 (s, 1H), 6.91 (d, J=2.3 Hz, 2H), 5.07 (s, 2H), 4.95 (s, 1H) ppm.

NUCC-200495: ¹H NMR (500 MHz, CDCl₃) δ 7.96 (d, J=1.7 Hz, 2H), 7.90 (s,1H), 7.50 (d, J=8.6 Hz, 1H), 6.97-6.90 (m, 1H), 6.60 (d, J=8.6 Hz, 1H)ppm.

NUCC-198407: ¹H NMR (500 MHz, CDCl₃) δ 7.45-7.33 (m, 4H), 7.13 (d, J=8.5Hz, 1H), 6.59 (d, J=8.6 Hz, 1H), 6.48 (s, 1H), 5.15 (s, 2H), 3.79 (s,3H) ppm.

NUCC-198399: ¹H NMR (500 MHz, CDCl₃) δ 7.79-7.72 (m, 2H), 7.68 (dt,J=7.8, 1.4 Hz, 1H), 7.66-7.59 (m, 1H), 7.35-7.32 (m, 2H), 7.24 (d, J=8.6Hz, 1H), 7.17 (t, J=6.9 Hz, 2H), 6.74 (t, J=5.7 Hz, 1H), 6.59 (s, 1H),5.07 (s, 2H), 3.87 (s, 3H) ppm.

NUCC-198398: ¹H NMR (500 MHz, CDCl₃) δ 7.78 (s, 1H), 7.75-7.66 (m, 2H),7.61 (t, J=8.5 Hz, 2H), 7.32 (d, J=8.5 Hz, 2H), 7.13 (t, J=12.8 Hz, 2H),6.93 (s, 1H), 6.72 (d, J=8.8 Hz, 1H), 5.06 (s, 2H) ppm.

NUCC-198394: ¹H NMR (500 MHz, CDCl₃) δ 8.00 (d, J=8.3 Hz, 1H), 7.62 (d,J=8.6 Hz, 1H), 7.55 (d, J=8.3 Hz, 1H), 7.38 (s, 1H), 7.14 (d, J=8.3 Hz,1H), 6.88 (s, 1H), 6.74 (d, J=8.9 Hz, 1H), 5.07 (s, 2H) ppm.

NUCC-198391: ¹H NMR (500 MHz, CDCl₃) δ 7.79 (t, J=8.4 Hz, 1H), 7.59 (d,J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 1H), 7.12 (t, J=10.8 Hz, 2H), 6.93 (s,1H), 6.76-6.65 (m, 1H), 5.06 (s, 2H) ppm.

NUCC-198363: ¹H NMR (500 MHz, CDCl₃) δ 7.64 (d, J=8.7 Hz, 2H), 7.50-7.43(m, 2H), 7.41-7.35 (m, 1H), 7.31-7.26 (m, 4H), 7.15 (d, J=8.5 Hz, 1H),6.86 (s, 1H), 5.07 (s, 2H) ppm.

NUCC-196362: ¹H NMR (500 MHz, CDCl₃) δ 7.94 (s, 2H), 7.85 (s, 1H), 7.58(d, J=8.7 Hz, 2H), 7.27 (d, J=2.0 Hz, 1H), 7.13 (d, J=8.3 Hz, 2H), 6.95(s, 1H), 6.70 (d, J=8.7 Hz, 1H), 5.03 (s, 2H) ppm.

NUCC-196361: ¹H NMR (500 MHz, CDCl₃) δ 7.88 (s, 2H), 7.84 (s, 1H),7.28-7.16 (m, 3H), 7.11 (d, J=8.1 Hz, 2H), 6.73 (d, J=8.5 Hz, 1H), 6.56(s, 1H), 5.01 (s, 2H), 3.80 (s, 3H) ppm; ¹³C NMR (126 MHz, CDCl₃) δ157.51, 151.53, 142.21, 141.91, 139.42, 134.56, 134.27, 134.05, 131.82,131.74, 131.47, 131.41, 131.21, 131.08, 129.16, 128.92, 128.82, 128.33,128.21, 124.42, 122.26, 122.19, 121.62, 120.05, 115.89, 115.77, 109.80,107.55, 105.51, 105.30, 69.88 ppm.

NUCC-196344: ¹H NMR (500 MHz, CDCl₃) δ 7.53 (d, J=8.7 Hz, 1H), 7.45 (d,J=8.3 Hz, 1H), 7.35 (d, J=7.7 Hz, 2H), 7.26 (t, J=6.9 Hz, 3H), 7.21 (d,J=7.7 Hz, 1H), 7.11 (d, J=8.3 Hz, 2H), 6.78 (s, 1H), 6.66 (d, J=8.8 Hz,1H), 5.02 (s, 2H), 2.43 (s, 3H) ppm.

C. Synthetic Method C

Synthetic Method C

Step 1.

Over a solution of 1-(2,4-dihydroxyphenyl)ethanone (0.92 g, 6.05 mmol)and pyridinium p-toluenesulfonate (0.061 g, 0.242 mmol) were introducedin 9 mL of dichloromethane, 3,6-dihydro-2H-pyran (1.655 ml, 18.14 mmol,1 equiv.) was added. Then, the resulting solution stirred at rt for 3 h.Reaction was quenched by adding 9 mL of an aqueous saturated solution ofNaHCO₃. Layers were separated and aqueous layer was extracted withdichloromethane (2×10) mL. Combined organic layers were dried overNa₂SO₄, filtrated and concentrated. After that, the resulting solid wassolved in 50 mL of EtOH and ethyl oxalate was added (18.15 mmol, 3equiv.). The resulting solution was added dropwise over a suspension ofsodium ethoxide (30.3 equiv.) in 10 mL of ethanol. After the addition,the reaction was heated at 90 C for 30 minutes. The reaction was cooleddown and 50 mL of DCM and 20 mL of HCl 3M were added. Layers wereseparated, and aqueous layer was extracted with dichloromethane (2×50mL). Combined organic layers were dried over Na₂SO₄, filtrated andconcentrated. The yellow solid obtained was then solved in 50 mL of amixture 1:1 of dichloromethane:THF and pTsOH (0.6 mmol, 1 equiv.) wereadded and it stirred at RT for 1.5 hours. The reaction was directlyconcentrated under reduced pressure. Finally, over a solution of thesolid obtained, iodine (24 mmol) in 75 mL of CHCl₃, pyridine (24 mmol, 4equiv.) were added. The resulting solution was stirred at roomtemperature for 16 h. Then, 75 mL of saturated aqueous Na₂S₂O₃ wereadded and the resulting mixture stirred for one hour. The organic layerwas separated and the aqueous phase was extracted with CH₂Cl₂ (3×60 mL).The combined organic layers were washed with brine and dried overanhydrous Na₂SO₄. Solvent was then removed under reduced pressure andthe residue was triturated with diethyl ether several times.

Step 2.

A suspension of 2-carbonyl-chromenone, (3 mmol), the halo-alkane (3.6mmol, 1.2 equiv.) and K₂CO₃ (6 mmol) in 30 mL of acetone was heated at60° C. for 16 h. The reaction was filtered through a funnel and thesolvent removed under reduced pressure. The crude residue was trituratedwith water and dried under reduced pressure until dryness.

Step 3.

A suspension of the previous alkylated chromenone (0.34 mmol, 1 equiv.),with the corresponding boronic acid (0.37 mmol, 1.1 equiv.), Na₂CO₃(0.68 mmol, 2 equiv.) and Pd(dppf)Cl₂ (0.026 mmol, 0.08 equiv.) in 3.5mL of a mixture 1:2:6 of EtOH:water:toluene was bubbled with nitrogengas for 10 minutes. Then, the flask was capped, and the mixture washeated at 90° C. for 2 h. The dark solution was cool down to roomtemperature and diluted with EtOAc. The organic layer was separated andthe aqueous phase was extracted with EtOAc (3×3 mL). The combinedorganic layers were washed with brine and dried over anhydrous Na₂SO₄.Solvent was then removed under reduced pressure and the residue waspurified by silica gel chromatography.

Step 4.

A solution of the previous chromenone (0.2 mmol, 1 equiv.) with thedesired hydrazine (0.6 mmol, 3 equiv.) in 2 mL of EtOH was heated at 70°C. for 45 minutes. The solution was cooled down to room temperature andconcentrated. The solid residue was directly purified by silica gelchromatography (n-hexanes/ethyl acetate=5:1 to 1:1) providing thedesired pyrazole.

Step 5.

Over a solution of the ethyl-esterpyrazole (0.1 mmol, 1 equiv.) in 0.5mL of a mixture THF:water 1:1, a drop of 10% NaOH aqueous solution wasadded stirred at RT for 24 h. The reaction was concentrated to dryness.Then, the resulting solid was solved in 1 mL of DMF and TSTU (0.1 mmol,1 equiv.) and DIPEA (0.2 mmol) were added and the solution stirred for20 minutes at RT. After that, the amine (0.2 mmol) were added andstirred at RT for 30 minutes. The reaction was directly purified bypreparative reverse phase HPLC.

Examples

NUCC-201224: ¹H NMR (500 MHz, CDCl₃) δ 11.00 (s, 1H), 7.96 (d, J=1.6 Hz,2H), 7.79 (s, 1H), 7.46 (d, J=8.7 Hz, 1H), 7.32-7.18 (m, 2H), 7.13 (d,J=8.4 Hz, 2H), 6.74 (s, 1H), 6.62 (d, J=8.7 Hz, 1H), 6.09 (q, J=4.9 Hz,1H), 4.99 (s, 2H), 4.14 (s, 3H), 2.99 (d, J=4.9 Hz, 3H) ppm.

NUCC-200973: ¹H NMR (500 MHz, CDCl₃) δ 7.90 (s, 2H), 7.72 (s, 1H), 7.52(d, J=8.6 Hz, 1H), 7.27 (s, 1H), 7.18 (d, J=8.0 Hz, 2H), 7.07 (d, J=8.5Hz, 2H), 7.03 (s, 1H), 6.60 (d, J=8.6 Hz, 1H), 4.95 (s, 2H) ppm.

NUCC-200558: ¹H NMR (500 MHz, CDCl₃) δ 10.90 (s, 1H), 7.77 (t, J=1.6 Hz,1H), 7.70 (dt, J=7.8, 1.6 Hz, 1H), 7.60 (dt, J=7.8, 1.6 Hz, 1H),7.53-7.43 (m, 2H), 7.27 (d, J=8.3 Hz, 2H), 7.12 (d, J=8.3 Hz, 2H), 6.81(s, 1H), 6.60 (d, J=8.7 Hz, 1H), 5.01 (s, 2H), 4.15 (s, 3H) ppm.

D. Synthetic Method D

Synthetic Method D

Step 1.

In an appropriate sized vial, substituted phenol (1 equiv.), aryl/alkylchloride (1.1 equiv.), K₂CO₃ (3 equiv.) in dry acetone were added andstirred at 60° C. overnight. On completion, the solvent was evaporatedand the residue was suspended in EtOAc (10 mL). The organic portion iswashed with H₂O (2×10 mL). The combined aqueous portion was furtherextracted with EtOAc (10 mL). The combined organic portion was washedwith brine, dried over Na₂SO₄ and evaporated to yield a crude residue.The residue was used for the next step with further purification.

Step 2.

In an appropriate sized vial,(1-methyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)boronic acid (1 equiv.),step 1 product (1 equiv.), Pd(dppf)₂Cl₂ (0.05 equiv.), Na₂CO₃ (3 equiv.)in a mixture of dioxane:H₂O (1:1). The vial were flushed with N₂ for 1min and stirred at 100° C. for 1 h. On completion the reaction mixturewas cooled to room temp and passed through a silica plug using DCM:MeOH(10:1). The solvent was evaporated to yield the crude residue. The crudewas purified by prep HPLC. To obtain the desired product. egNUCC-0200813: ¹H NMR (500 MHz, CDCl₃) δ 7.32 (s, 1H), 7.28-7.19 (m, 4H),7.12 (s, 1H), 7.05 (d, J=8.4 Hz, 1H), 7.01 (d, J=2.8 Hz, 1H), 6.77 (dd,J=8.5, 2.8 Hz, 1H), 4.94 (s, 2H), 4.34 (s, 2H), 3.86 (s, 3H); ¹³C NMR(126 MHz, CDCl₃) δ 158.7, 140.9, 135.3, 133.9, 132.5, 131.9, 128.8,121.7, 118.9, 114.3, 113.7, 69.3, 63.3, 39.6.

Step 3.

In an appropriate sized vial, add step 2 product (1 equiv.) and NaBH₄(1.2 equiv.) in methanol and stirred at room temp for 7 h. Oncompletion, the reaction was concentrated and suspended in H₂O. Thesuspension was extracted with EtOAc (3×3 mL) and the combined organicportion was evaporated to yield crude residue which was purified by prepHPLC

Example

NUCC-0200813: prep HPLC condition (40-80%, 5 min, 50×30 mm, Rt=3.74min). ¹H NMR (500 MHz, CDCl₃) δ 7.32 (s, 1H), 7.30-7.18 (m, 4H),7.10-6.95 (m, 2H), 6.77 (dd, J=8.5, 2.8 Hz, 1H), 4.94 (s, 2H), 4.34 (s,2H), 3.86 (s, 3H) ppm; ¹³C NMR (126 MHz, CDCl₃) δ 158.7, 140.9, 135.3,133.8, 132.5, 131.9, 128.8, 121.73, 118.9, 114.3, 113.7, 69.3, 63.3,39.6 ppm.

Step 4.

In an appropriate sized vial, add product of step 2 (1 equiv.), NBS (5equiv.) in anh. DMF and stir the reagents at room temp for 3 days. Oncompletion, the reaction was quenched with water (4 mL) and extractedwith EtOAc (3×3 mL). The combined organic portion was dried and theresidue was purified via Biotage (5:1 Hex/EtOAc; 10 g column). Collectedfractions: 11-14. eg NUCC-0200979: ¹H NMR (500 MHz, CDCl₃) δ 7.42-7.34(m, 6H), 7.31 (dd, J=8.7, 2.6 Hz, 1H), 5.11 (s, 2H), 3.82 (s, 3H); ¹³CNMR (126 MHz, CDCl₃) δ 159.7, 140.2, 140.1, 139.8, 134.6, 133.6, 133.3,129.1, 128.8, 123.1, 120.1, 119.4, 119.3, 116.3, 115.1, 93.7, 70.0,38.8.

Step 5.

In an appropriate sized vial, add product of step 2 (1 equiv.), H₂O₂(30% aqueous solution, 1 equiv.), Na₂CO₃ (3 equiv.) in MeOH was stirredat room temp overnight. On completion the solvent was evaporated and theresidue was suspended in water (1 mL) and extracted with EtOAc (3×1 mL).The combined organic portion was evaporated to yield the crude which waspurified by prep HPLC. eg NUCC-0200816: ¹H NMR (500 MHz, CDCl₃) δ 7.33(d, J=2.7 Hz, 1H), 7.24 (s, 4H), 7.19-7.04 (m, 2H), 6.99 (dd, J=8.5, 2.7Hz, 1H), 6.40 (s, 1H), 5.47 (s, 1H), 5.23 (s, 1H), 4.98 (s, 2H), 3.58(s, 3H); ¹³C NMR (126 MHz, CDCl₃) δ 168.6, 159.7, 142.6, 136.6, 134.4,134.2, 132.8, 128.9, 128.8, 119.3, 117.8, 115.1, 104.9, 69.6, 37.5.

E. Synthetic Method E

Synthetic Method E

Step 1.

Over a solution of 1-(2,4-dihydroxyphenyl)ethanone (0.92 g, 6.05 mmol, 1equiv.) and PYRIDINIUM P-TOLUENESULFONATE (0.061 g, 0.242 mmol, 0.04equiv.) were introduced in 9 mL of dichloromethane, 3,6-dihydro-2H-pyran(1.655 ml, 18.14 mmol, 3 equiv.) was added. Then, the resulting solutionstirred at rt for 3 h. Reaction was quenched by adding 9 mL of anaqueous saturated solution of NaHCO₃. Layers were separated and aqueouslayer was extracted with dichloromethane (2×10) mL. Combined organiclayers were dried over The redish oil-solid was then solved in DMF-DMA(9.1 mmol) and was heated at 95° C. for 3 h. Then, it was cool down andconcentrated under reduced pressure. The solid obtained was then solvedin 50 mL of a mixture 1:1 of dichloromethane:THF and pTsOH (0.6 mmol, 1equiv.) were added and it stirred at RT for 1.5 hours. The reaction wasdirectly concentrated under reduced pressure. Finally, over a solutionof the solid obtained, iodine (24 mmol, 4 equiv.) in 75 mL of CHCl₃,pyridine (24 mmol, 4 equiv.) were added. The resulting solution wasstirred at room temperature for 16 h. Then, 75 mL of saturated aqueousNa₂S₂O₃ were added and the resulting mixture stirred for one hour. Theorganic layer was separated and the aqueous phase was extracted withCH₂Cl₂ (3×60 mL). The combined organic layers were washed with brine anddried over anhydrous Na₂SO₄. Solvent was then removed under reducedpressure and the residue was triturated with diethyl ether severaltimes.

Step 2.

A suspension of the chromenone, (3 mmol, v), the halo-alkane (3.6 mmol,1.2 equiv.) and K₂CO₃ (6 mmol) in 30 mL of acetone was heated at 60° C.for 16 h. The reaction was filtered through a funnel and the solventremoved under reduced pressure. The crude residue was triturated withwater and dried under reduced pressure until dryness.

Step 3.

A suspension of the previous alkylated chromenone (0.34 mmol, 1 equiv.),with the corresponding boronic acid (0.37 mmol, 1.1 equiv.), Na₂CO₃(0.68 mmol, 2 equiv.) and Pd(dppf)Cl₂ (0.026 mmol, 0.08 equiv.) in 3.5mL of a mixture 1:2:6 of EtOH:water:toluene was bubbled with nitrogengas for 10 minutes. Then, the flask was capped, and the mixture washeated at 90° C. for 2 h. The dark solution was cool down to roomtemperature and diluted with EtOAc. The organic layer was separated andthe aqueous phase was extracted with EtOAc (3×3 mL). The combinedorganic layers were washed with brine and dried over anhydrous Na₂SO₄.Solvent was then removed under reduced pressure and the residue waspurified by silica gel chromatography.

Step 4.

A solution of the previous chromenone (0.2 mmol, 1 equiv.) with thedesired hydrazine (0.6 mmol, 3 equiv.) in 2 mL of EtOH was heated at 70°C. for 45 minutes. The solution was cooled down to room temperature andconcentrated. The solid residue was directly purified by silica gelchromatography (n-hexanes/ethyl acetate=5:1 to 1:1) providing thedesired pyrazole.

Examples

NUCC-198359: ¹H NMR (500 MHz, CDCl₃) δ 7.45 (s, 1H), 7.35 (s, 3H), 7.00(s, 2H), 6.82 (d, J=8.8 Hz, 1H), 6.62 (d, J=2.6 Hz, 1H), 6.22 (dd,J=8.8, 2.6 Hz, 1H), 4.99 (s, 3H), 2.38 (s, 3H), 2.03 (d, J=6.1 Hz, 6H)ppm.

NUCC-198322: ¹H NMR (500 MHz, CDCl₃) δ 7.80 (s, 1H), 7.51 (dt, J=9.0,4.5 Hz, 1H), 7.40-7.37 (m, 4H), 7.32 (d, J=7.3 Hz, 1H), 6.66 (d, J=2.6Hz, 1H), 6.52-6.41 (m, 3H), 5.05 (s, 2H) ppm.

NUCC-198318: ¹H NMR (500 MHz, CDCl₃) δ 7.87 (s, 1H), 7.85 (s, 2H), 7.77(s, 1H), 7.38 (s, 4H), 6.95 (d, J=8.7 Hz, 1H), 6.69 (d, J=2.6 Hz, 1H),6.34 (dd, J=8.7, 2.6 Hz, 1H), 5.04 (s, 2H).

NUCC-196350: ¹H NMR (500 MHz, CDCl₃) δ 7.61 (s, 1H), 7.45 (s, 1H), 7.36(s, 1H), 7.33 (s, 3H), 7.31-7.28 (m, 2H), 7.08-7.01 (m, 2H), 6.62 (d,J=2.6 Hz, 1H), 6.29 (dd, J=8.7, 2.6 Hz, 1H), 4.99 (s, 2H) ppm.

NUCC-196348: ¹H NMR (500 MHz, CDCl₃) δ 7.41-7.33 (m, 2H), 7.31-7.06 (m,7H), 6.66 (d, J=8.6 Hz, 1H), 6.51 (s, 1H), 5.02 (s, 2H), 3.82 (s, 3H)ppm.

F. Synthetic Method F

Synthetic Method F

Step 1.

Over a mixture of the 2-phenylacetic acid derivative (10.58 mmol, 1equiv.) and resorcinol (1.165 g, 10.58 mmol, 1 equiv.), Borontrifluoride etherate ORON (4.02 ml, 31.7 mmol, 3 equiv.) was added. Thevial was sealed and it stirred at 90° C. for 1.5 h. The reaction wascool down to RT. The solid obtained was partitioned between 20 mL ofwater and 20 mL of dichloromethane. Layers were separated and theaqueous one was extracted with dichloromethane 3×10 mL). The combinedorganic layers were dried over Na₂SO₄ filtrated and concentrated underreduced pressure. Thee residue was purified by silica gelchromatography.

Step 2.

A suspension of the O-phenoxy-acetophenone (1 mmol) with TFFA (5 equiv)and pyridine (5 equiv.) was heated at 120° C. for 4 h. Then, it was cooldown to r.t. In some cases product was precipitated, in others water wasadded and then extracted with EtOAc (3×10 mL). The combined organiclayers were washed with brine, dried over Na₂SO₄, filtrated andconcentrated. The residue was purified by silica gel chromatography.

Step 3.

A suspension of the-chromenone, (0.5 mmol, 1 equiv.), K₂CO₃ (1 mmol) andthe halo alkane (0.6 mmol) in 5 mL of acetone was heated at 60° C. for16 h. The reaction was filtered through a funnel and the solvent removedunder reduced pressure. The crude residue was triturated with water anddried under reduced pressure until dryness.

Step 4.

A solution of the previous O-alkylated-chromenone (0.2 mmol) with thedesired hydrazine (0.6 mmol, 3 equiv.) in 2 mL of EtOH was heated at 70°C. for 45 minutes. The solution was cooled down to room temperature andconcentrated. The solid residue was directly purified by silica gelchromatography (n-hexanes/ethyl acetate=5:1 to 1:1) providing thedesired pyrazole.

Examples

NUCC-198309: ¹H NMR (500 MHz, CDCl₃) δ 7.38-7.33 (m, 5H), 7.00-6.84 (m,4H), 6.55 (s, 1H), 6.35 (s, 1H), 5.00 (s, 2H), 3.81 (s, 3H) ppm.

NUCC-198295: ¹H NMR (500 MHz, CDCl₃) δ 7.44 (d, J=2.2 Hz, 3H), 7.38-7.31(m, 6H), 6.89 (s, 1H), 6.55 (s, 1H), 6.32 (s, 1H), 5.00 (s, 2H) ppm.

NUCC-196340: ¹H NMR (500 MHz, CDCl₃) δ 7.97 (s, 1H), 7.45 (d, J=9.0 Hz,1H), 7.37-7.28 (m, 2H), 7.18 (d, J=8.7 Hz, 2H), 6.86 (d, J=8.7 Hz, 2H),6.45 (d, J=2.6 Hz, 1H), 6.37 (dd, J=9.0, 2.6 Hz, 1H), 5.92 (s, 1H), 5.01(s, 2H), 3.78 (s, 3H) ppm.

NUCC-196282: ¹H NMR (500 MHz, CDCl₃) δ 7.34-7.28 (m, 4H), 6.94 (s, 1H),6.92-6.84 (m, 2H), 6.79 (d, J=1.8 Hz, 1H), 6.52 (s, 1H), 6.31 (s, 1H),4.97 (s, 2H), 3.92 (s, 3H), 3.78 (s, 3H) p.

G. Synthetic Method G

Synthetic Method G

Step 1.

Over a suspension of 1-(2,4-dihydroxyphenyl)ethan-1-one (5.00 g, 32.89mmol, 1 equiv.) in trifluoroacetic anhydride (18.50 mL, 131.56 mmol, 4equiv.) placed in a high-pressure tube, sodium 2,2,2-trifluoroacetate(9.84 g, 72.36 mmol, 2.2 equiv.) was added and the system was capped andstirred at 110° C. for 24 h. The reaction was allowed to cool down toapproximately 70° C. and then was diluted with 200 mL of EtOAc. Themixture was neutralized by adding saturated aqueous K₂CO₃ solution untilno more bubbling was observed. Layers were separated and the aqueousphase was extracted with more EtOAc (3×150 mL). The combined organiclayers were washed with brine and dried over anhydrous Na₂SO₄. Thesolution was then concentrated to 100-150 mL of EtOAc. Then the flaskwas capped and kept at room temperature for 1-2 days, obtaining a solidwhich was filtrated and dried under vacuum to obtain 4.09 g of pure 1 asa white solid in 54% yield.

Then, over an ice-bath solution of the solid obtained (2 g, 8.70 mmol, 1equiv.) and pyridine (2.81 mL, 34.8 mmol, 4 equiv.) in 18 mL of DCMunder nitrogen atmosphere, Tf₂O (2.20 mL, 13.06 mmol, 1.5 equiv.) wasadded dropwise for 15 min. Then, the mixture stirred from 0° C. to roomtemperature for 16 h. The reaction was quenched by adding 15 mL ofwater. The organic layer was separated and the aqueous one was extractedwith EtOAc (3×15 mL). The combined organic layers were washed with brineand dried over anhydrous Na₂SO₄. Solvent was then removed under reducedpressure and the residue was purified by silica gel chromatography(n-hexanes/ethyl acetate=10:1 to 4:1) providing compound 2 as a yellowsolid in 95% yield (3.0 g): mp 46-48° C. ¹H NMR (500 MHz, DMSO-d₆) δ8.32-8.20 (m, 2H), 7.74 (dd, J=8.8, 2.4 Hz, 1H), 7.17 (s, 1H) ppm. ¹³CNMR (126 MHz, DMSO-d₆) δ 175.4, 155.6, 152.4, 151.4 (q, ²J (C,F)=38.8Hz), 128.2, 123.7, 120.1, 118.5 (q, ¹J (C,F)=274.7 Hz), 112.9, 111.5,111.4 ppm. HRMS (ESI): mass calc for C₁₁H₅F₆O₅S⁺ [M+H]⁺=362.9756,found=362.9758.

Step 2.

In order to obtain different Pd-catalyzed coupling products in theposition 7 of the 2-CF₃-chromenone, different conditions were followed:

Conditions A: Aniline Coupling:

Over a suspension of the chromenone-triflate (150 mg, 0.41 mmol, 1equiv.), with Cs₂CO₃ (202 mg, 0.62 mmol, 1 equiv.), BINAP (25 mg, 0.04mmol, 0.1 equiv.) and PdOAc₂ (4.5 mg, 0.02 mmol, 0.05 equiv.) in 4 mL oftoluene was bubbled with nitrogen gas for 10 minutes. Then, the flaskwas capped, and the mixture was heated at 90° C. for 16 h. The darksolution was cool down to room temperature and diluted with 5 mL ofEtOAc and 5 mL of water. The organic layer was separated and the aqueousphase was extracted with EtOAc₂ (3×5 mL). The combined organic layerswere washed with brine and dried over anhydrous Na₂SO₄. Solvent was thenremoved under reduced pressure and the residue was purified by silicagel chromatography.

Conditions B: Phenol Coupling:

Over a suspension of the chromenone-triflate (150 mg, 0.41 mmol, 1equiv.), with K₂CO₃ (113 mg, 0.82 mmol, 2 equiv.), JohnPhos (12 mg, 0.04mmol, 0.1 equiv.) and Pd₂dba₃ (19 mg, 0.02 mmol, 0.05 equiv.) in 4 mL oftoluene, nitrogen gas was bubbled for 10 minutes. Then, the flask wascapped, and the mixture was heated at 90° C. for 16 h. The dark solutionwas cool down to room temperature and diluted with 5 mL of EtOAc and 5mL of water. The organic layer was separated and the aqueous phase wasextracted with EtOAc₂ (3×5 mL). The combined organic layers were washedwith brine and dried over anhydrous Na₂SO₄. Solvent was then removedunder reduced pressure and the residue was purified by silica gelchromatography.

Conditions C: Boronic Acid Coupling to Biaryl Chromenones:

A suspension of the chromenone-triflate (150 mg, 0.41 mmol, 1 equiv.),with the corresponding boronic acid (0.46 mmol, 1.1 equiv.), Na₂CO₃ (87mg, 0.82 mmol, 2 equiv.) and Pd(dppf)Cl₂ (23 mg, 0.03 mmol, 0.08 equiv.)in 4 mL of a mixture 1:2:6 of EtOH:water:toluene was bubbled withnitrogen gas for 10 minutes. Then, the flask was capped, and the mixturewas heated at 90° C. for 20 min. The dark solution was cool down to roomtemperature and diluted with EtOAc. The organic layer was separated andthe aqueous one was extracted with EtOAc₂ (3×3 mL). The combined organiclayers were washed with brine and dried over anhydrous Na₂SO₄. Solventwas then removed under reduced pressure and the residue was purified bysilica gel chromatography.

Step 3.

A solution of the previous 7-functionalized chromenone (0.2 mmol, 1equiv.) with the desired hydrazine (0.6 mmol, 3 equiv.) in 2 mL of EtOHwas heated at 70° C. for 45 minutes. The solution was cooled down toroom temperature and concentrated. The solid residue was directlypurified by silica gel chromatography (n-hexanes/ethyl acetate=5:1 to1:1) providing the desired pyrazole.

Examples

NUCC-201223: ¹H NMR (500 MHz, CDCl₃) δ 10.62 (s, 1H), 7.92 (d, J=1.7 Hz,2H), 7.81 (s, 1H), 7.56 (d, J=8.7 Hz, 1H), 7.23-7.19 (m, 2H), 6.98 (s,1H), 6.86-6.77 (m, 2H), 6.61 (d, J=8.7 Hz, 1H) ppm.

NUCC-200721: ¹H NMR (500 MHz, CDCl₃) δ 7.46 (d, J=8.4 Hz, 1H), 7.31 (d,J=8.7 Hz, 1H), 7.20 (d, J=2.7 Hz, 1H), 6.92 (dd, J=8.7, 2.7 Hz, 1H),6.84 (s, 1H), 6.62 (d, J=7.7 Hz, 2H), 5.78 (s, 1H) ppm.

NUCC-200681: ¹H NMR (500 MHz, CDCl₃) δ 7.72 (d, J=8.8 Hz, 1H), 7.28 (d,J=8.8 Hz, 2H), 7.07 (d, J=8.5 Hz, 2H), 6.94 (d, J=8.8 Hz, 2H), 6.81 (d,J=8.7 Hz, 1H), 6.52 (d, J=2.4 Hz, 1H), 6.45 (d, J=8.8 Hz, 1H), 2.57 (q,J=7.6 Hz, 2H), 1.18 (t, J=7.6 Hz, 3H) ppm.

NUCC-200679: ¹H NMR (500 MHz, CDCl₃) δ 7.51 (d, J=8.6 Hz, 1H), 7.33-7.28(m, 2H), 7.01-6.97 (m, 2H), 6.88 (s, 1H), 6.60 (dd, J=8.5, 2.4 Hz, 1H),6.56 (d, J=2.4 Hz, 1H) ppm.

NUCC-200559: ¹H NMR (500 MHz, CDCl₃) δ 7.58 (d, J=8.7 Hz, 1H), 7.41 (d,J=8.4 Hz, 1H), 7.05 (q, J=9.1 Hz, 1H), 6.96-6.90 (m, 2H), 6.87 (dd,J=8.7, 2.4 Hz, 1H), 6.77 (d, J=8.2 Hz, 2H), 6.56 (dd, J=8.5, 2.2 Hz,1H), 6.52 (d, J=2.2 Hz, 1H), 5.72 (s, 1H) ppm.

NUCC-200492: ¹H NMR (500 MHz, CDCl₃) δ 7.28-7.25 (m, 2H), 7.09-7.06 (m,2H), 7.06-7.02 (m, 1H), 6.61 (d, J=7.7 Hz, 2H), 6.53 (s, 1H), 3.81 (s,3H) ppm.

NUCC-200491: ¹H NMR (500 MHz, CDCl₃) δ 7.46 (d, J=9.0 Hz, 1H), 7.25 (d,J=8.7 Hz, 2H), 7.10-6.97 (m, 2H), 6.81 (s, 1H), 6.62 (dq, J=5.3, 2.2 Hz,2H), 5.80 (s, 1H) ppm.

H. Synthetic Method H

Synthetic Method H

Step 1.

Over a suspension of 1-(2,4-dihydroxyphenyl)ethan-1-one (5.00 g, 32.89mmol, 1 equiv.) in trifluoroacetic anhydride (18.50 mL, 131.56 mmol, 4equiv.) placed in a high-pressure tube, sodium 2,2,2-trifluoroacetate(9.84 g, 72.36 mmol, 2.2 equiv.) was added and the system was capped andstirred at 110° C. for 24 h. The reaction was allowed to cool down toapproximately 70° C. and then was diluted with 200 mL of EtOAc. Themixture was neutralized by adding saturated aqueous K₂CO₃ solution untilno more bubbling was observed. Layers were separated and the aqueousphase was extracted with more EtOAc (3×150 mL). The combined organiclayers were washed with brine and dried over anhydrous Na₂SO₄. Thesolution was then concentrated to 100-150 mL of EtOAc. Then the flaskwas capped and kept at room temperature for 1-2 days, obtaining a solidwhich was filtrated and dried under vacuum to obtain 4.09 g of pure 1 asa white solid in 54% yield.

Step 2.

A suspension of 7-hydroxy-2-(trifluoromethyl)-4H-chromen-4-one, (1 g,2.8 mmol, 1 equiv.), the halo-alkane (3.4 mmol, 1.2 equiv.) and K₂CO₃(0.77 g, 5.6 mmol, 2 equiv.) in 5 mL of acetone was heated at 60° C. for16 h. The reaction was filtered through a funnel and the solvent removedunder reduced pressure. The crude residue was triturated with water anddried under reduced pressure until dryness.

Step 3.

Synthesis of pyrazoles. A solution of the previous chromenone (0.2 mmol,1 equiv.) with the desired hydrazine (0.6 mmol, 3 equiv.) in 2 mL ofEtOH was heated at 70° C. for 45 minutes. The solution was cooled downto room temperature and concentrated. The solid residue was directlypurified by silica gel chromatography (n-hexanes/ethyl acetate=5:1 to1:1) providing the desired pyrazole.

Example

NUCC-198314: ¹H NMR (500 MHz, CDCl₃) δ 7.12 (s, 1H), 6.97 (d, J=8.5 Hz,1H), 6.51 (dd, J=8.5, 2.4 Hz, 1H), 6.46 (d, J=2.4 Hz, 1H), 6.41 (s, 1H),4.92 (s, 2H), 3.67 (s, 3H) ppm.

Synthesis of Pyrimidines.

A solution of the previous chromenone (0.2 mmol, 1 equiv.) with thedesired benzimidamide (0.26 mmol, 1.3 equiv.) and potassium hydroxide(0.6 mmol, 3 equiv.) in 10 mL of EtOH was heated at 80° C. for 14 hours.The solution was diluted in 5 mL of water, extracted 3×15 mL of EtOAc.Combined organic layers were dried over Na₂SO₄, filtrated andconcentrated under reduced pressure. The solid residue was directlypurified by silica gel chromatography (n-hexanes/ethyl acetate=5:1 to1:1) providing the desired pyrimidine.

Example

NUCC-0200500: ¹H NMR (500 MHz, CDCl₃) δ 8.80-8.73 (m, 2H), 8.07 (s, 1H),7.95 (s, 1H), 7.81 (d, J=8.9 Hz, 1H), 7.37 (s, 4H), 6.69-6.59 (m, 2H),5.10 (s, 2H) ppm.

I. Synthetic Method I

Synthetic method I (PAL)

Steps 1-4.

Same as described in Synthetic method B above.

Step 5.

In a 100 mL round bottom flask, dimethyl malonate (1.1 equiv.),propargyl bromide (1 equiv.), K₂CO₃ (3 equiv.) in dry acetone wasstirred at room temp for 36 h. The reaction mixture was quenched byaddition of sat. NH₄Cl solution and extracted with DCM (3 times). Thecombined organic layers were washed with H₂O (2 times), dried overNa₂SO₄ and the solvent was evaporated under reduced pressure to yield ayellow-colored oil which converted into a solid on standing. (Note:Conversion could be monitored by TLC: H:EtOAc:5:1, rf=0.42). The crudewas stirred with KOH (1 equiv.) in MeOH at room temp for 4. Oncompletion, the solvent was evaporated and the residue was suspended inH₂O and washed with Et₂O (2 times). The aqueous portion was acidifiedwith HCl (2N, to pH 3) and extracted with EtOAc (2 times). The organicportion was dried over Na₂SO₄ and evaporated to yield the crudeyellow-colored oil. (Note: LCMS: shows m/z=157 and 174 (+18)).

Step 6.

In a appropriate sized vial, product from step 5 (1 equiv.), IBX-SO3K(1.5 equiv.), NaI (0.2 equiv.), NaN₃ (3.3 equiv.) in anhydrous DMSO werestirred in an ice-cold water bath for 5 min and further heated at 60° C.for 2 h. On completion, the reaction mixture was quenched with sat.Na₂S₂O₃ (25 mL) and extracted with Et₂O (2×25 mL). The combined organicportion was washed with sat. NaHCO₃ (2×25 mL) and dried over Na₂SO₄. Theorganic portion was evaporated to yield a yellow-colored residue. Thecrude along with LiOH (2 equiv.) in THF:H₂O (1:1) was stirred at roomtemp for 2 h. (Note: LCMS shows a dimer signal for the SM and productindicating that reaction is complete). The solvent was evaporated andthe crude was taken for the next step without any further purification.

Step 7.

In an appropriate sized vial, product from step 4 (1 equiv.), productfrom step 5 (1.5 equiv.), HATU (1.5 equiv.), DIEA (4 equiv.) inanhydrous DCM was stirred at 30° C. for 6 h. On completion, the solventwas evaporated to yield a crude residue. The crude was purified by prepHPLC.

Examples

NUCC_0201698: Prep HPLC: (25-90%, 50×30, C18, 50 mL/min, Rt=4.3 min).UVmax=222 nm ¹H NMR (500 MHz, CDCl₃) δ 8.84 (dd, J=10.0, 1.9 Hz, 3H),8.16 (s, 1H), 8.02 (s, 2H), 7.50 (d, J=8.5 Hz, 2H), 7.24 (s, 17H), 6.76(d, J=8.6 Hz, 1H), 6.59 (s, 2H), 5.22 (s, 1H), 5.06 (s, 2H), 4.28 (dd,J=7.6, 4.1 Hz, 1H), 3.83 (s, 3H), 3.09-2.75 (m, 3H), 2.23-2.06 (m, 1H)ppm.

NUCC-0201694: Prep HPLC: (25-90%, 50×30, C18, 50 mL/min, Rt=3.5 min).UVmax=222 nm ¹H NMR (500 MHz, CDCl₃) δ 10.63 (s, 1H), 7.80 (d, J=2.0 Hz,1H), 7.70-7.37 (m, 3H), 6.91 (s, 1H), 6.58 (d, J=8.7 Hz, 1H), 6.41 (s,1H), 4.13-3.85 (m, 4H), 3.27 (dq, J=34.6, 6.8 Hz, 1H), 2.99-2.57 (m,2H), 1.94-1.77 (m, 1H), 1.23 (s, 1H), 0.81 (s, 1H) ppm.

NUCC-0201695: Prep HPLC: (25-90%, 50×30, C18, 50 mL/min, Rt=4.5-5 min).UVmax=222 nm ¹H NMR (500 MHz, CDCl₃) δ 7.76 (d, J=2.0 Hz, 1H), 7.68-7.48(m, 2H), 7.19 (d, J=8.6 Hz, 1H), 6.65 (d, J=8.7 Hz, 1H), 6.55 (s, 1H),6.43 (s, 1H), 5.09 (s, 1H), 4.04 (dt, J=30.2, 6.5 Hz, 3H), 3.82 (s, 3H),3.27 (dt, J=36.6, 6.8 Hz, 2H), 2.96-2.82 (m, 1H), 2.77-2.62 (m, 1H),2.05 (t, J=2.7 Hz, 1H), 1.98-1.81 (m, 2H) ppm.

NUCC-0201696: Prep HPLC: (25-90%, 50×30, C18, 50 mL/min, Rt=4.6 min).UVmax=222 nm ¹H NMR (500 MHz, CDCl₃) δ 8.85 (dd, J=9.8, 2.1 Hz, 3H),8.06 (d, J=2.2 Hz, 1H), 7.24 (s, 13H), 6.69 (d, J=8.6 Hz, 1H), 6.59 (s,1H), 6.45 (s, 1H), 5.27 (s, 1H), 4.19-3.93 (m, 3H), 3.84 (s, 3H), 3.28(ddd, J=47.0, 13.7, 6.8 Hz, 2H), 2.88 (ddd, J=17.2, 4.3, 2.6 Hz, 1H),2.69 (ddd, J=17.1, 7.2, 2.7 Hz, 1H), 1.90 (dt, J=7.7, 3.9 Hz, 2H) ppm.

NUCC-0210697: Prep HPLC: (25-90%, 50×30, C18, 50 mL/min, Rt=3.8-4.1min). UVmax=222 nm ¹H NMR (500 MHz, CDCl₃) δ 9.03-8.72 (m, 2H), 8.13 (d,J=11.1 Hz, 1H), 7.69-7.40 (m, 1H), 6.95 (d, J=5.2 Hz, 1H), 6.69-6.43 (m,2H), 4.07 (ddd, J=11.4, 8.8, 5.1 Hz, 3H), 3.71-3.41 (m, 2H), 2.95-2.59(m, 3H), 2.45 (dq, J=4.9, 2.4 Hz, 1H), 2.04-1.88 (m, 1H) ppm.

J. Synthetic Method J

Synthetic Method J (PROTAC)

Steps 1-4.

Same as described in Synthetic method above.

Step 5.

In an appropriate sized vial, the azido-PEG carboxylicacid (1 equiv.)was stirred with thionyl chloride (39 equiv.) at room temp for 3 h. Oncompletion, (LCMS shows the methyl ester in indicating reactioncompletion) the excess thionyl chloride was evaporated to yield thecrude acid chloride. A solution of product from step 4 (1 equiv.) wasadded in anhydrous THE was added to the acid chloride and TEA (5 equiv.)and the reaction mixture was stirred at 60° C. for 3 h. On completionthe mixture was filtered through a cotton plug and purified by prep HPLC(50×30, C18, 50 mL/min, Rt. 3.3-3.6 min) which was taken up for thecoupling step without further purification (assuming quantitativeyield).

Step 6.

In an appropriate sized vial, product from step 5 (1 equiv.), CUSO₄ (5equiv.), sodium ascorbate (5 equiv.), propargyl-CRBN (1 equiv.) inTHF:H₂O (1:1) was stirred at room temp overnight. On completion, thereaction mixture was diluted with ACN (1 mL) and purified by prep HPLC.

Examples

NUCC-0201202: Prep HPLC (45-95% 50×30, C18, 50 mL/min, Rt=3.2-3.7 min).UVmax=222 nm. ¹H NMR (500 MHz, CDCl₃) δ 8.80 (s, 1H), 8.26 (s, 1H), 7.90(s, 2H), 7.79 (s, 1H), 7.71 (s, 1H), 7.65-7.43 (m, 5H), 7.37 (t, J=7.9Hz, 2H), 7.13 (d, J=8.1 Hz, 2H), 6.86 (s, 1H), 6.68 (d, J=8.7 Hz, 1H),5.26 (s, 2H), 5.04-4.79 (m, 3H), 4.32 (s, 2H), 4.08 (s, 2H), 3.84-3.45(m, 13H), 3.04-2.51 (m, 5H), 2.23-1.95 (m, 4H), 1.57 (s, 4H) ppm.

NUCC-0201203: Prep HPLC (45-95% 50×30, C18, 50 mL/min, Rt=3.25-4 min).UVmax=222 nm. ¹H NMR (500 MHz, CDCl₃) δ 8.80 (s, 1H), 8.29 (s, 1H), 7.91(s, 3H), 7.78 (d, J=11.5 Hz, 3H), 7.61 (t, J=7.9 Hz, 2H), 7.52 (d, J=8.3Hz, 4H), 7.42 (dd, J=14.9, 7.9 Hz, 3H), 7.14 (d, J=8.1 Hz, 2H), 6.87 (s,1H), 6.68 (d, J=8.7 Hz, 1H), 5.31 (s, 2H), 5.07-4.79 (m, 4H), 4.37 (t,J=5.1 Hz, 2H), 4.07 (s, 3H), 3.85-3.32 (m, 20H), 3.01-2.53 (m, 5H),2.26-1.93 (m, 4H), 1.58 (s, 4H) ppm.

NUCC_0201660: Prep HPLC (45-95% 50×30, C18, 50 mL/min, Rt=4.25 min).UVmax=222 nm.

NUCC-0201702: ¹H NMR (500 MHz, CDCl₃) δ 8.35 (s, 1H), 7.86 (s, 1H), 7.75(d, J=2.0 Hz, 1H), 7.67 (dd, J=8.5, 7.3 Hz, 1H), 7.61-7.37 (m, 4H), 7.17(d, J=8.6 Hz, 1H), 6.96 (t, J=6.2 Hz, 1H), 6.64 (d, J=8.6 Hz, 1H), 6.53(s, 1H), 5.39 (d, J=2.8 Hz, 3H), 4.89 (dd, J=12.3, 5.4 Hz, 1H),4.60-4.40 (m, 2H), 4.08-3.90 (m, 3H), 3.81 (d, J=5.3 Hz, 4H), 3.69-3.48(m, 7H), 3.27 (q, J=6.8 Hz, 2H), 2.26-2.03 (m, 4H), 1.85 (q, J=6.4 Hz,2H), 1.23 (d, J=2.2 Hz, 3H) ppm.

NUCC-0201703: Prep HPLC (20-80% 50×30, C18, 50 mL/min, Rt=4.15 min).Uvmax=222 nm. Unable to open NMR-fid

NUCC-0201704: Prep HPLC (20-80% 50×30, C18, 50 mL/min, Rt=4 min).Uvmax=222 nm. ¹H NMR (500 MHz, CDCl₃) δ 8.82 (d, J=33.9 Hz, 2H), 8.07(s, 1H), 7.97 (s, 1H), 7.77 (s, 1H), 7.63 (t, J=7.9 Hz, 1H), 7.53 (d,J=8.8 Hz, 1H), 7.48-7.37 (m, 1H), 7.02 (d, J=4.2 Hz, 1H), 6.88 (s, 1H),6.61 (d, J=8.8 Hz, 1H), 5.28 (d, J=3.0 Hz, 1H), 4.92 (dd, J=11.9, 5.5Hz, 1H), 4.40 (t, J=5.1 Hz, 1H), 4.07 (t, J=5.1 Hz, 1H), 3.94 (s, 1H),3.75 (t, J=5.1 Hz, 1H), 3.68-3.41 (m, 4H), 3.02-2.64 (m, 2H), 2.15 (s,1H), 1.24 (s, 2H), 0.84 (d, J=22.7 Hz, 1H) ppm.

In vitro metabolism of NUCC-176242 and NUCC-176248 were tested usingmouse liver microsomes and a mouse S9 fraction. NUCC-176242 wassignificantly metabolism by the mouse S9 fraction versus NUCC-176248likely due to S9 conjugation at the N-1 nitrogen atom of the pyrazolering.

The pharmacokinetics of NUCC-176242 and NUCC-176248 were studied in miceby administering a dose of 5 mg/kg intravenously and measuring theplasma concentration versus time. The observed in vivo metabolism ofNUCC-176242 and NUCC-176248 correlated well with the observed in vitrometabolism tested above for of NUCC-176242 and NUCC-176248.

Tables

TABLE 1 Representative Compounds Molecular Molecule weight SynthesisName (g/mol) Structure Method NUCC- 0176234 452.5

— NUCC- 0176242 488.9

A NUCC- 0176243 420.9

A NUCC- 0176244 454.4

A NUCC- 0176245 536.5

A NUCC- 0176246 390.4

A NUCC- 0176247 392.8

A NUCC- 0176248 502.9

A NUCC- 0176249 376.4

A NUCC- 0176250 455.4

A NUCC- 0176251 359.4

A NUCC- 0176252 373.4

A NUCC- 0176253 406.9

A NUCC- 0176254 387.4

A NUCC- 0176255 358.4

A NUCC- 0176256 392.5

A NUCC- 0176257 404.4

A NUCC- 0176258 418.5

A NUCC- 0176259 390.4

A NUCC- 0176260 434.9

A NUCC- 0176261 434.9

A NUCC- 0176262 420.9

A NUCC- 0196282 449.0

A NUCC- 0196283 504.9

F NUCC- 0196284 420.9

A NUCC- 0196285 434.9

A NUCC- 0196286 420.9

A NUCC- 0196287 434.9

A NUCC- 0196288 416.5

A NUCC- 0196289 430.5

A NUCC- 0196290 436.5

A NUCC- 0196291 450.5

A NUCC- 0196294 463.0

A NUCC- 0196295 488.9

A NUCC- 0196296 502.9

A NUCC- 0196297 484.5

A NUCC- 0196298 498.5

A NUCC- 0196299 421.5

A NUCC- 0196301 478.8

A NUCC- 0196302 492.9

A NUCC- 0196303 427.3

A NUCC- 0196304 509.3

A NUCC- 0196305 441.3

F NUCC- 0196306 505.9

A NUCC- 0196311 518.9

F NUCC- 0196312 474.9

F NUCC- 0196313 368.7

H NUCC- 0196314 382.8

H NUCC- 0196340 488.9

F NUCC- 0196341 497.9

A NUCC- 0196342 495.3

A NUCC- 0196343 427.3

A NUCC- 0196344 458.9

B NUCC- 0196345 390.9

E NUCC- 0196346 404.9

E NUCC- 0196347 462.8

B NUCC- 0196348 476.9

B NUCC- 0196349 408.9

E NUCC- 0196350 411.3

E NUCC- 0196351 425.3

E NUCC- 0196352 517.0

A NUCC- 0196353 486.9

A NUCC- 0196354 537.4

A NUCC- 0196355 523.3

A NUCC- 0196356 517.0

A NUCC- 0196357 502.9

A NUCC- 0196358 498.5

A NUCC- 0196359 484.5

A NUCC- 0196360 594.9

B NUCC- 0196361 594.9

B NUCC- 0196362 580.8

B NUCC- 0196363 479.3

B NUCC- 0196364 459.9

B NUCC- 0196365 459.9

B NUCC- 0196366 445.8

B NUCC- 0198293 376.8

F NUCC- 0198294 390.9

F NUCC- 0198295 444.8

F NUCC- 0198296 458.9

F NUCC- 0198297 458.9

F NUCC- 0198298 390.9

F NUCC- 0198299 404.9

F NUCC- 0198300 404.9

F NUCC- 0198301 404.9

F NUCC- 0198302 418.9

F NUCC- 0198303 418.9

F NUCC- 0198304 406.9

F NUCC- 0198305 420.9

F NUCC- 0198306 434.9

F NUCC- 0198307 434.9

F NUCC- 0198308 406.9

F NUCC- 0198309 474.9

F NUCC- 0198310 488.9

F NUCC- 0198311 488.9

F NUCC- 0198312 420.9

F NUCC- 0198313 434.9

F NUCC- 0198314 434.9

F NUCC- 0198315 434.9

F NUCC- 0198316 449.0

F NUCC- 0198317 449.0

F NUCC- 0198318 512.8

E NUCC- 0198319 526.9

E NUCC- 0198320 377.8

E NUCC- 0198321 391.9

E NUCC- 0198322 366.8

F NUCC- 0198323 380.8

E NUCC- 0198324 380.8

E NUCC- 0198325 434.8

B NUCC- 0198326 420.9

E NUCC- 0198352 404.9

E NUCC- 0198353 418.9

E NUCC- 0198354 418.9

E NUCC- 0198355 390.9

E NUCC- 0198356 404.9

E NUCC- 0198357 406.9

E NUCC- 0198358 420.9

E NUCC- 0198359 418.9

E NUCC- 0198360 433.0

E NUCC- 0198361 380.8

E NUCC- 0198362 394.9

E NUCC- 0198391 469.9

B NUCC- 0198392 483.9

B NUCC- 0198393 483.9

B NUCC- 0198394 487.9

B NUCC- 0198395 514.9

B NUCC- 0198396 529.0

B NUCC- 0198397 529.0

B NUCC- 0198398 469.9

B NUCC- 0198399 483.9

B NUCC- 0198400 483.9

B NUCC- 0198401 487.9

B NUCC- 0198402 501.9

B NUCC- 0198403 514.9

B NUCC- 0198404 529.0

B NUCC- 0198405 529.0

B NUCC- 0198406 364.3

A NUCC- 0198407 461.7

B NUCC- 0198408 461.7

B NUCC- 0198409 501.9

B NUCC- 0198410 542.9

B NUCC- 0198411 378.4

A NUCC- 0198412 382.8

H NUCC- 0200489 473.9

C NUCC- 0200490 493.3

— NUCC- 0200491 353.7

G NUCC- 0200492 367.8

G NUCC- 0200493 367.8

G NUCC- 0200494 381.8

G NUCC- 0200495 456.3

B NUCC- 0200496 380.3

F NUCC- 0200497 243.2

G NUCC- 0200498 257.2

G NUCC- 0200499 458.9

H NUCC- 0200500 524.9

H NUCC- 0200501 592.9

H NUCC- 0200502 395.8

H NUCC- 0200503 474.8

H NUCC- 0200557 444.9

B NUCC- 0200558 458.9

B NUCC- 0200559 355.3

G NUCC- 0200560 369.3

G NUCC- 0200561 363.3

G NUCC- 0200562 377.3

G NUCC- 0200563 362.3

G NUCC- 0200564 376.3

G NUCC- 0200565 320.3

G NUCC- 0200566 334.3

G NUCC- 0200567 326.3

G NUCC- 0200568 340.4

G NUCC- 0200569 385.4

G NUCC- 0200570 368.7

D NUCC- 0200571 469.9

D NUCC- 0200572 458.9

D NUCC- 0200573 368.7

D NUCC- 0200574 228.2

H NUCC- 0200575 462.8

D NUCC- 0200576 479.3

D NUCC- 0200577 442.8

NUCC- 0200677 328.8

G NUCC- 0200678 310.7

G NUCC- 0200679 354.7

G NUCC- 0200680 473.9

G NUCC- 0200681 474.9

G NUCC- 0200682 377.4

A NUCC- 0200683 348.3

A NUCC- 0200684 362.4

A NUCC- 0200685 238.3

— NUCC- 0200686 502.9

B NUCC- 0200687 243.2

D NUCC- 0200688 242.2

D NUCC- 0200689 244.2

D NUCC- 0200690 367.8

D NUCC- 0200691 366.8

D NUCC- 0200692 368.7

D NUCC- 0200721 388.2

G NUCC- 0200722 327.4

— NUCC- 0200723 470.3

B NUCC- 0200724 470.3

B NUCC- 0200725 512.8

B NUCC- 0200726 526.9

B NUCC- 0200727 526.9

B NUCC- 0200728 547.3

B NUCC- 0200729 561.3

B NUCC- 0200730 512.8

B NUCC- 0200731 526.9

B NUCC- 0200732 547.3

B NUCC- 0200733 561.3

B NUCC- 0200734 530.8

B NUCC- 0200735 544.9

B NUCC- 0200736 544.9

B NUCC- 0200737 547.3

B NUCC- 0200738 561.3

B NUCC- 0200739 396.8

D NUCC- 0200740 410.8

D NUCC- 0200741 367.8

D NUCC- 0200742 366.8

D NUCC- 0200743 384.8

D NUCC- 0200744 401.2

D NUCC- 0200745 381.8

D NUCC- 0200746 381.8

D NUCC- 0200747 368.7

D NUCC- 0196314.2 382.8

H NUCC- 0198412.2 382.8

H NUCC- 0200812 391.8

D NUCC- 0200813 396.8

D NUCC- 0200814 401.2

D NUCC- 0200815 394.8

D NUCC- 0200816 409.8

D NUCC- 0200817 395.8

D NUCC- 0200970 584.9

C NUCC- 0200971 570.9

C NUCC- 0200972 584.9

C NUCC- 0200973 556.9

C NUCC- 0200974 526.9

B NUCC- 0200975 561.3

B NUCC- 0200976 394.8

D NUCC- 0200977 384.8

D NUCC- 0200978 396.8

D NUCC- 0200979 470.7

— NUCC- 0201023 561.4

B NUCC- 0201024 594.9

B NUCC- 0201025 608.9

B NUCC- 0201026 608.9

B NUCC- 0201027 590.4

B NUCC- 0201028 604.5

B NUCC- 0201029 604.5

B NUCC- 0201030 508.4

B NUCC- 0201031 512.4

B NUCC- 0201032 553.4

B NUCC- 0201033 564.4

B NUCC- 0201034 578.4

B NUCC- 0201035 578.4

B NUCC- 0201036 522.9

B NUCC- 0201037 513.8

B NUCC- 0201038 470.8

B NUCC- 0201039 557.9

B NUCC- 0201040 463.8

B NUCC- 0201041 576.4

B NUCC- 0201042 590.4

B NUCC- 0201043 590.4

B NUCC- 0201192 473.4

B NUCC- 0201193 402.4

B NUCC- 0201194 430.4

B NUCC- 0201195 416.4

B NUCC- 0201196 540.4

B NUCC- 0201197 498.3

B NUCC- 0201198 526.4

B NUCC- 0201199 776.6

J NUCC- 0201200 820.7

J NUCC- 0201201 561.4

B NUCC- 0201202 1088.9

J NUCC- 0201203 1133.0

J NUCC- 0201204 459.4

B NUCC- 0201205 487.4

B NUCC- 0201206 473.4

B NUCC- 0201207 444.5

B NUCC- 0201208 430.4

B NUCC- 0201209 459.4

B NUCC- 0201210 487.4

B NUCC- 0201211 473.4

B NUCC- 0201212 444.5

B NUCC- 0201213 430.4

B NUCC- 0201219 600.4

B NUCC- 0201220 470.3

B NUCC- 0201221 484.3

B NUCC- 0201222 484.3

B NUCC- 0201223 566.8

G NUCC- 0201224 583.9

C NUCC- 0201225 478.8

B NUCC- 0201226 484.9

B NUCC- 0201227 492.9

B NUCC- 0201630 527.9

B NUCC- 0201631 527.9

B NUCC- 0201632 484.9

B NUCC- 0201633 484.9

B NUCC- 0201634 492.9

B NUCC- 0201635 506.9

B NUCC- 0201636 459.4

B NUCC- 0201637 445.4

B NUCC- 0201638 416.4

B NUCC- 0201639 416.4

B NUCC- 0201640 479.8

B NUCC- 0201641 479.8

B NUCC- 0201642 493.8

B NUCC- 0201643 493.8

B NUCC- 0201644 532.4

B NUCC- 0201645 432.3

NUCC- 0201646 479.8

B NUCC- 0201647 465.8

B NUCC- 0201648 605.4

B NUCC- 0201649 605.4

B NUCC- 0201650 600.9

NUCC- 0201651 526.4

B NUCC- 0201652 526.4

B NUCC- 0201653 538.4

B NUCC- 0201654 538.4

B NUCC- 0201655 560.5

B NUCC- 0201656 560.5

B NUCC- 0201657 594.0

B NUCC- 0201658 594.0

B NUCC- 0201659 559.9

I NUCC- 0201660 1000.9

J NUCC- 0201661 434

B NUCC- 0201694 581.5

I NUCC- 0201695 614.9

I NUCC- 0201696 581.5

I NUCC- 0201697 553.4

I NUCC- 0201698 629.5

I NUCC- 0201699 629.5

I NUCC- 0201702 1021.3

J NUCC- 0201703 987.9

J NUCC- 0201704 959.8

J NUCC- 0201705 312.3

J NUCC- 0201909 450.9

B NUCC- 0201910 408.8

B NUCC- 0201911 537.9

B NUCC- 0201912 551.9

B NUCC- 0201913 551.9

B

TABLE 2 Biological Activity of Representative Compounds Potency code <10++++ 10-30 +++ 30-40 ++ EMSA Molecule Name >40 + IC50(uM) NUCC-0176234 +123.1 NUCC-0176242 ++ 89 NUCC-0176243 + NUCC-0176244 + NUCC-0176245 ++95 NUCC-0176246 + NUCC-0176247 + NUCC-0176248 + 96 NUCC-0176249 +NUCC-0176250 + NUCC-0176251 + NUCC-0176252 + NUCC-0176253 +NUCC-0176254 + NUCC-0176255 + NUCC-0176256 + NUCC-0176257 +NUCC-0176258 + NUCC-0176259 + NUCC-0176260 + NUCC-0176261 +NUCC-0176262 + NUCC-0196282 + NUCC-0196283 ++++ 49 NUCC-0196284 +NUCC-0196285 + NUCC-0196286 + NUCC-0196287 + NUCC-0196288 +NUCC-0196289 + NUCC-0196290 + NUCC-0196291 + NUCC-0196294 + NUCC-0196295+++ 66.8 NUCC-0196296 + NUCC-0196297 + NUCC-0196298 ++ NUCC-0196299 +NUCC-0196301 + NUCC-0196302 + NUCC-0196303 + NUCC-0196304 +++NUCC-0196305 ++++ NUCC-0196306 + NUCC-0196311 + NUCC-0196312 +NUCC-0196313 ++++ NUCC-0196314 + NUCC-0196340 ++ NUCC-0196341 +NUCC-0196342 ++++ 64.4 NUCC-0196343 + NUCC-0196344 +++ NUCC-0196345 +NUCC-0196346 + NUCC-0196347 + NUCC-0196348 +++ NUCC-0196349 +NUCC-0196350 +++ NUCC-0196351 + NUCC-0196352 + NUCC-0196353 +NUCC-0196354 + NUCC-0196355 +++ NUCC-0196356 + NUCC-0196357 +++NUCC-0196358 + NUCC-0196359 + NUCC-0196360 + NUCC-0196361 ++++ 63NUCC-0196362 ++ 102.2 NUCC-0196363 ++++ 61.8 NUCC-0196364 +NUCC-0196365 + NUCC-0196366 + NUCC-0198293 + NUCC-0198294 + NUCC-0198295++++ 53.3 NUCC-0198296 + NUCC-0198297 + NUCC-0198298 + NUCC-0198299 +NUCC-0198300 + NUCC-0198301 + NUCC-0198302 + NUCC-0198303 +NUCC-0198304 + NUCC-0198305 + NUCC-0198306 + NUCC-0198307 +NUCC-0198308 + NUCC-0198309 ++++ 61.2 NUCC-0198310 + NUCC-0198311 +NUCC-0198312 + NUCC-0198313 + NUCC-0198314 + NUCC-0198315 +NUCC-0198316 + NUCC-0198317 + NUCC-0198318 ++ NUCC-0198319 +NUCC-0198320 + NUCC-0198321 + NUCC-0198322 ++ NUCC-0198323 +NUCC-0198324 + NUCC-0198325 + NUCC-0198326 + NUCC-0198352 +NUCC-0198353 + NUCC-0198354 + NUCC-0198355 + NUCC-0198356 +NUCC-0198357 + NUCC-0198358 + NUCC-0198359 ++ NUCC-0198360 +NUCC-0198361 + NUCC-0198362 + NUCC-0198391 ++++ NUCC-0198392 +NUCC-0198393 + NUCC-0198394 ++ NUCC-0198395 + NUCC-0198396 +NUCC-0198397 + NUCC-0198398 ++++ NUCC-0198399 ++++ NUCC-0198400 +NUCC-0198401 + NUCC-0198402 + NUCC-0198403 + NUCC-0198404 +NUCC-0198405 + NUCC-0198406 ++++ NUCC-0198407 +++ NUCC-0198408 +NUCC-0198409 + NUCC-0198410 + NUCC-0198411 +++ NUCC-0198412 +NUCC-0200489 + NUCC-0200490 + NUCC-0200491 ++++ NUCC-0200492 +++NUCC-0200493 + NUCC-0200494 + NUCC-0200495 ++++ NUCC-0200496 +NUCC-0200497 + NUCC-0200498 + NUCC-0200499 + NUCC-0200500 +NUCC-0200501 + NUCC-0200502 + NUCC-0200503 + NUCC-0200557 + NUCC-0200558++++ NUCC-0200559 ++ NUCC-0200560 + NUCC-0200561 + NUCC-0200562 +NUCC-0200563 + NUCC-0200564 + NUCC-0200565 + NUCC-0200566 +NUCC-0200567 + NUCC-0200568 + NUCC-0200569 + NUCC-0200570 +NUCC-0200571 + NUCC-0200572 + NUCC-0200573 + NUCC-0200574 + NUCC-0200575++++ NUCC-0200576 ++++ NUCC-0200677 + NUCC-0200678 + NUCC-0200679 ++NUCC-0200680 + NUCC-0200681 ++++ NUCC-0200682 + NUCC-0200683 +++NUCC-0200684 + NUCC-0200685 + NUCC-0200686 + NUCC-0200687 +NUCC-0200688 + NUCC-0200689 + NUCC-0200690 + NUCC-0200691 +NUCC-0200692 + NUCC-0200721 ++++ NUCC-0200722 + NUCC-0200723 ++NUCC-0200724 + NUCC-0200725 + NUCC-0200726 + NUCC-0200727 +NUCC-0200728 + NUCC-0200729 + NUCC-0200730 + NUCC-0200731 +NUCC-0200732 + NUCC-0200733 + NUCC-0200734 + NUCC-0200735 +NUCC-0200736 + NUCC-0200737 + NUCC-0200738 +++ NUCC-0200739 +NUCC-0200740 + NUCC-0200741 + NUCC-0200742 + NUCC-0200743 +NUCC-0200744 + NUCC-0200745 + NUCC-0200746 + NUCC-0200747 +NUCC-0196314.2 + NUCC-0198412.2 + NUCC-0200812 + NUCC-0200813 +NUCC-0200814 + NUCC-0200815 + NUCC-0200816 + NUCC-0200817 +NUCC-0200970 + NUCC-0200971 ++++ NUCC-0200972 + NUCC-0200973 ++++NUCC-0200974 + NUCC-0200975 + NUCC-0200976 + NUCC-0200977 +NUCC-0200978 + NUCC-0200979 + NUCC-0201023 + NUCC-0201024 ++++NUCC-0201025 ++ NUCC-0201026 + NUCC-0201027 +++ NUCC-0201028 +NUCC-0201029 + NUCC-0201030 + NUCC-0201031 + NUCC-0201032 +NUCC-0201033 + NUCC-0201034 + NUCC-0201035 + NUCC-0201036 ++NUCC-0201037 +++ NUCC-0201038 ++++ NUCC-0201039 +++ NUCC-0201040 +++NUCC-0201041 + NUCC-0201042 + NUCC-0201043 + NUCC-0201192 +++NUCC-0201193 ++++ NUCC-0201194 + NUCC-0201195 ++++ NUCC-0201196 +++NUCC-0201197 +++ NUCC-0201198 +++ NUCC-0201199 +++ NUCC-0201200 +NUCC-0201201 ++++ NUCC-0201202 +++ NUCC-0201203 ++ NUCC-0201204 +++NUCC-0201205 ++++ NUCC-0201206 ++++ NUCC-0201207 +++ NUCC-0201208 ++++NUCC-0201209 + NUCC-0201210 + NUCC-0201211 + NUCC-0201212 + NUCC-0201213++++ NUCC-0201219 ++++ NUCC-0201220 ++++ NUCC-0201221 + NUCC-0201222 +++NUCC-0201223 ++++ NUCC-0201224 ++++ NUCC-0201225 + NUCC-0201226 +++NUCC-0201227 ++++ NUCC-0201630 + NUCC-0201631 + NUCC-0201632 +++NUCC-0201633 + NUCC-0201634 ++ NUCC-0201635 + NUCC-0201636 +NUCC-0201637 + NUCC-0201638 + NUCC-0201639 + NUCC-0201640 +NUCC-0201641 + NUCC-0201642 + NUCC-0201643 + NUCC-0201644 +NUCC-0201646 + NUCC-0201647 + NUCC-0201648 + NUCC-0201649 +NUCC-0201651 + NUCC-0201652 + NUCC-0201653 +++ NUCC-0201654 +NUCC-0201655 + NUCC-0201656 + NUCC-0201657 + NUCC-0201658 ++NUCC-0201659 + NUCC-0201660 + NUCC-0201661 + NUCC-0201694 +NUCC-0201695 + NUCC-0201696 ++ NUCC-0201697 + NUCC-0201698 +NUCC-0201699 + NUCC-0201702 + NUCC-0201703 + NUCC-0201704 +NUCC-0201705 + NUCC-0201909 + NUCC-0201910 + NUCC-0201911 +NUCC-0201912 + NUCC-0201913 +

TABLE 3 Biological Activity of Representative Compounds Plasma ProteinMYCCAP MLM % Binding Molecule Name IC50(uM)^(a) PC12 IC50 ^(b) at 60min^(c) (mouse) NUCC-0176242 3.1 9.8 NUCC-0176245 4.1 NUCC-0176248 3.47.8 NUCC-0196283 2.3 NUCC-0196295 12.2 NUCC-0196312 4.8 NUCC-0196313 4.2NUCC-0196342 3.5 87.4 NUCC-0196344 6.6 NUCC-0196348 110.0 NUCC-01963504.3 NUCC-0196355 11.8 NUCC-0196357 6.9 NUCC-0196360 39.6 NUCC-01963613.8 30.7 72.9 >99.9% NUCC-0196362 4.0 NUCC-0196363 4.0 88.6 NUCC-01982954.6 NUCC-0198309 4.0 74 NUCC-0198318 4.3 NUCC-0198322 6.5 NUCC-01983597.6 NUCC-0198391 3.4 NUCC-0198394 7.3 NUCC-0198398 4.3 NUCC-0198399 2.580.3 NUCC-0198406 17.5 NUCC-0198407 7.8 NUCC-0200558 49.3 31.0NUCC-0200739 2.2 19.4 46.4 NUCC-0200972 5.1 81.0 60.3 >99.9%NUCC-0200973 68.3 29.6 NUCC-0200974 10.2 NUCC-0200975 6.1 38.4NUCC-0200976 3.9 63.3 21.8 >99.9% NUCC-0201032 1.0 11.0 30.5 >99.9%NUCC-0201038 6.8 18.3 NUCC-0201039 13.6 NUCC-0201041 12.6 127.7NUCC-0201194 4.2 59.8 NUCC-0201195 14.9 70.3 NUCC-0201196 4.5 66.6NUCC-0201197 3.4 18.6 NUCC-0201198 2.6 8.2 NUCC-0201199 3.0 12.7NUCC-0201202 4.0 23.1 NUCC-0201203 32.4 NUCC-0201204 23.0 NUCC-02012059.5 36.3 NUCC-0201206 5.0 NUCC-0201207 5.7 52.7 NUCC-0201209 4.4 10.3NUCC-0201212 23.7 30.8 NUCC-0201219 9.5 NUCC-0201220 4.9 91.6NUCC-0201221 1.2 NUCC-0201223 5.1 40.6 75.8 NUCC-0201224 1.1 11.6NUCC-0201225 19.0 82.6 NUCC-0201227 4.5 1.0 NUCC-0201630 15.4 42.3NUCC-0201633 4.6 27.3 NUCC-0201656 5.5 18.0 NUCC-0201699 24.3 ^(a)Cellviability of MycCap cells ^(b) Cell viability of PC12 cells ^(c)%remaining of compound after 60 min treatment with mouse liver microsomes

REFERENCES

-   [1] Huang M, Weiss W A. 2013. Neuroblastoma and MYNC. Cold Spring    Harb Perspect Med 3: a014415.-   [2] Roussel M F, Robinson G W. 2013. Role of MYC in medulloblastoma.    Cold Spring Harb Perspect Med 3: a014308.-   [3] Gabay M, Li Y, Felsher D W. 2014. MYC activation is a hall mark    of cancer initiation and maintenance. Cold Spring Harb Perspect Med    doi: 10.1101/cshperspect.a014241.-   [4] Schmitz R, Ceribelli M, et. al. 2014. Oncogenic mechanisms in    Burkitt lymphoma. Cold Spring Harb Perspect Med 4: a014282.-   [5] Michael R. McKeown and James E. Bradner, Cold Spring Harb    Perspect Med 2014; 4:a014266-   [6] Soucek L, Whitfield J R, et. al. 2013. Inhibition of MYC family    proteins eradicates KRas-driven lung cancer in mice. Genes Dev 27:    504-513.-   [7] S. Fletcher, E. V. Prochownik, Small-molecule inhibitors of the    MYC oncoprotein, Biochim. Biophys. Acta (2014).

Example 2—Biological Assays

Proliferation Assay (FIG. 11).

PC3 prostate cancer cell line with high Myc level and PC12pheochromocytoma tumor cell line with non functional Max protein, whichis not dependent on Myc-Max complex, were phased from ATCC. Cells wereplated in 96 well plate at 1000 cells per well, and Myc inhibitor 361 atvarious concentrations was added to the cells next day. After 3 days oftreatment, fresh medium with 361 were added again, and cell viabilitywas measured by MTS assay at day 5 after the treatment. (See FIG. 11).

Myc Ebox Luciferase Reporter Assay (FIG. 12).

MycCap cells stably expressing luciferase with CMV promoter (MycCap-luc)or c-Myc E-box-luciferase reporter (MycCap Ebox-luc) were plated at10000 cells per well in 96 well white-wall plate. Serial dilutions of361 were treated next day. At 4 hours of treatment, luminescence signalwas determined immediately after adding 150 μg/ml of Luciferin to thewell. (See FIG. 12).

Cellular Thermal Shift Assay (CETSA) (FIG. 13).

PC3 cells with 70 to 80% confluence in 15 cm culture dish were treatedwith 6 μM of 361 or vehicle (DMSO) for 30 min. Cells were harvested andwashed once with PBS, then suspended in 1 mL of PBS supplemented withproteinase and phosphatase inhibitors. The PBS contained 6 μM of 361 orvehicle (DMSO) accordingly at this step. The cell suspension wasdistributed into seven to ten 0.2-ml PCR tubes with 100 μl volume (about1 million cells) and each tube was designated a temperature point.Samples were heated at their designated temperatures for 2 min in AB96-well thermal cycler. Immediately after heating, remove and incubatethe tubes at room temperature for 3 min. After this 3 min incubation,immediately snap-freeze in liquid nitrogen, and stored at −80 C°. Tolysis cells, three freeze and thaw cycles in LN was performed. The tubesare vortexed briefly after each thawing. Cell lysis was collected andcell debris together with precipitated and aggregated proteins wereremoved by centrifuging samples at 20,000 g for 20 min at 4° C. Celllysis samples were boiled for 5 min at 90° C. after adding loadingbuffer, and ready for Western Blot analysis. c-Myc antibody is fromAbcam (Ab32072). Data was generated from three independent experiments,and c-Myc protein intensity was quantified through Image1. (See FIG.13).

Gene Expression Profiling Analysis (FIG. 14).

PC3 cells were treated with 10 μM of Min9-S1 for 24 hours. mRNA wasextracted using RNAeasy Plus mini kit (Qiagen, Cat. 74134). The geneexpression profiling was analyzed using HTA 2.0 from Affymetrix. GSEA offour Myc-dependent gene signature sets (Zeller et al., 2003; Schuhmacheret al., 2001; Kim et al., 2006; Schlosser et al., 2005) intranscriptional profiles of PC3 treated with Min9-S1 or vehicle (VEH)shows strong correlation with downregulation of expression by Min9-S1treatment. Gene sets suppressed in Min9-S1 treated PC3 cells includingthe number of genes in each set (n), the normalized enrichment score(NES), and test of statistical significance (FDR q value) were listed inthe table. (See FIG. 14).

MycCap FVB Allograft Model (FIG. 15).

FVB mice were inoculated with 1×10⁶ of MycCap cells in 100 ul ofmatrigel subcutaneously on both flanks of the mice. When tumor sizereached average size of 150 mm³, mice were randomized based on tumorvolume to 2 groups. Mice were administered with 50 mg/kg of 361 orvehicle(Veh) twice daily intra-peritoneally for two days. The treatmentwas suspended for 10 day, and initiated the treatment with a lower dose70 mg/kg daily for another 9 days. Tumor size was measured twice a weekduring the experiment. (See FIG. 15).

Myc Inhibitor 361 Combination with Immunotherapy (FIG. 16).

MycCap FVB allograft model was treated with 361 (50 mg/kg) for two days,following by two days of anti-PD-1 antibody (100 ug/day), and kept this4-day treatment cycle for 4 cycles. (See FIG. 16).

Myc Inhibitor Combination Treatment with Ara-C in AML Xenograft Model(FIG. 17).

CB17 SCID mice were inoculated with MV411 cells at the density of 5×10⁶suspended in PBS and matrigel (1:1). The mice were randomized based ontumor volume into 6 different groups after the tumor reached ˜200 to 500mm³. The mice were treated either with vehicle, NU031, NU975 alone or incombination with cytarabine (Ara-C). The combination treatment showssignificant difference compared to control group. (See FIG. 17).

NCI60 Profiling—NCI 60 Cell One-Dose Screen

General Description.

As of early 2007 all compounds submitted to the NCI 60 Cell screen aretested initially at a single high dose (10-5 M) in the full NCI 60 cellpanel. Only compounds which satisfy pre-determined threshold inhibitioncriteria in a minimum number of cell lines will progress to the full5-dose assay. The threshold inhibition criteria for progression to the5-dose screen was selected to efficiently capture compounds withanti-proliferative activity based on careful analysis of historical DTPscreening data. The threshold criteria may be updated as additional databecomes available.

Interpretation of One-Dose Data.

The One-dose data will be reported as a mean graph of the percent growthof treated cells and will be similar in appearance to mean graphs fromthe 5-dose assay. The number reported for the One-dose assay is growthrelative to the no-drug control, and relative to the time zero number ofcells. This allows detection of both growth inhibition (values between 0and 100) and lethality (values less than 0). This is the same as for the5-dose assay, described below. For example, a value of 100 means nogrowth inhibition. A value of 40 would mean 60% growth inhibition. Avalue of 0 means no net growth over the course of the experiment. Avalue of −40 would mean 40% lethality. A value of −100 means all cellsare dead. Information from the One-dose mean graph is available forCOMPARE analysis.

NCI 60 Cell Five-Dose Screen (FIG. 18, FIG. 19, and FIG. 20).

Compounds which exhibit significant growth inhibition in the One-DoseScreen are evaluated against the 60 cell panel at five concentrationlevels.

The human tumor cell lines of the cancer screening panel are grown inRPMI 1640 medium containing 5% fetal bovine serum and 2 mM L-glutamine.For a typical screening experiment, cells are inoculated into 96 wellmicrotiter plates in 100 μL at plating densities ranging from 5,000 to40,000 cells/well depending on the doubling time of individual celllines. After cell inoculation, the microtiter plates are incubated at37° C., 5% CO2, 95% air and 100% relative humidity for 24 h prior toaddition of experimental drugs.

After 24 h, two plates of each cell line are fixed in situ with TCA, torepresent a measurement of the cell population for each cell line at thetime of drug addition (Tz). Experimental drugs are solubilized indimethyl sulfoxide at 400-fold the desired final maximum testconcentration and stored frozen prior to use. At the time of drugaddition, an aliquot of frozen concentrate is thawed and diluted totwice the desired final maximum test concentration with complete mediumcontaining 50 μg/ml gentamicin. Additional four, 10-fold or ½ log serialdilutions are made to provide a total of five drug concentrations pluscontrol. Aliquots of 100 μl of these different drug dilutions are addedto the appropriate microtiter wells already containing 100 μl of medium,resulting in the required final drug concentrations.

Following drug addition, the plates are incubated for an additional 48 hat 37° C., 5% CO2, 95% air, and 100% relative humidity. For adherentcells, the assay is terminated by the addition of cold TCA. Cells arefixed in situ by the gentle addition of 50 μl of cold 50% (w/v) TCA(final concentration, 10% TCA) and incubated for 60 minutes at 4° C. Thesupernatant is discarded, and the plates are washed five times with tapwater and air dried. Sulforhodamine B (SRB) solution (100 μl) at 0.4%(w/v) in 1% acetic acid is added to each well, and plates are incubatedfor 10 minutes at room temperature. After staining, unbound dye isremoved by washing five times with 1% acetic acid and the plates are airdried. Bound stain is subsequently solubilized with 10 mM trizma base,and the absorbance is read on an automated plate reader at a wavelengthof 515 nm. For suspension cells, the methodology is the same except thatthe assay is terminated by fixing settled cells at the bottom of thewells by gently adding 50 μl of 80% TCA (final concentration, 16% TCA).Using the seven absorbance measurements [time zero, (Tz), controlgrowth, (C), and test growth in the presence of drug at the fiveconcentration levels (Ti)], the percentage growth is calculated at eachof the drug concentrations levels. Percentage growth inhibition iscalculated as:

[(Ti−Tz)/(C−Tz)]×100 for concentrations for which Ti>/=Tz

[(Ti−Tz)/Tz]×100 for concentrations for which Ti<Tz.

Three dose response parameters are calculated for each experimentalagent. Growth inhibition of 50% (GI50) is calculated from[(Ti−Tz)/(C−Tz)]×100=50, which is the drug concentration resulting in a50% reduction in the net protein increase (as measured by SRB staining)in control cells during the drug incubation. The drug concentrationresulting in total growth inhibition (TGI) is calculated from Ti=Tz. TheLC50 (concentration of drug resulting in a 50% reduction in the measuredprotein at the end of the drug treatment as compared to that at thebeginning) indicating a net loss of cells following treatment iscalculated from [(Ti−Tz)/Tz]×100=−50. Values are calculated for each ofthese three parameters if the level of activity is reached; however, ifthe effect is not reached or is exceeded, the value for that parameteris expressed as greater or less than the maximum or minimumconcentration tested. Results are shown in FIG. 18, FIG. 19, and FIG.20.

Example 3

Additional substituted heterocycles were synthesized and tested usingthe methods disclosed herein as indicated in the following Table.

TABLE Ebox reporter: IC50 CMV (uM); *** < 5 uM: ** reporter: IC50 5-10uM; * 10-50 Molecule Name Structure (uM) uM; NT-not tested NUCC-0202414

>50 ** NUCC-0202416

>50 *** NUCC-0202417

>50 >50 NUCC-0202418

>50 ** NUCC-0202421

>50 >50 NUCC-0202422

>50 >50 NUCC-0202423

>50 *** NUCC-0202424

>50 >50 NUCC-0202425

>50 * NUCC-0202426

>50 >50 NUCC-0202427

>50 * NUCC-0202428

>50 >50 NUCC-0202429

>50 *** NUCC-0202430

>50 >50 NUCC-0202431

39 * NUCC-0202432

28 * NUCC-0202433

>50 ** NUCC-0202434

40 *** NUCC-0202435

>50 * NUCC-0202436

>50 * NUCC-0202437

>50 >50 NUCC-0202521

34 * NUCC-0202522

43 * NUCC-0202523

>50 >50 NUCC-0202524

>50 * NUCC-0202525

>50 * NUCC-0202526

>50 * NUCC-0202527

>50 * NUCC-0202528

28 * NUCC-0202529

>50 * NUCC-0202530

>50 * NUCC-0202531

>50 >50 NUCC-0202532

>50 * NUCC-0202533

>50 * NUCC-0202534

>50 >50 NUCC-0202535

>50 * NUCC-0202536

44 ** NUCC-0202537

40 ** NUCC-0202538

>50 >50 NUCC-0202635

>50 * NUCC-0202636

41 ** NUCC-0202637

43 ** NUCC-0202638

>50 * NUCC-0202639

32 ** NUCC-0202640

>50 >50 NUCC-0202641

>50 >50 NUCC-0202642

>50 >50 NUCC-0202643

>50 *** NUCC-0202644

>50 * NUCC-0202648

>50 *** NUCC-0202649

>50 *** NUCC-0202650

>50 >50 NUCC-0202651

>50 ** NUCC-0202652

>50 >50 NUCC-0202686

31 *** NUCC-0202687

>50 *** NUCC-0202688

>50 ** NUCC-0202689

>50 >50 NUCC-0202690

>50 *** NUCC-0202691

>50 >50 NUCC-0202692

>50 ** NUCC-0202693

>50 *** NUCC-0202694

>50 >50 NUCC-0202695

>50 >50 NUCC-0202696

>50 >50 NUCC-0202697

>50 >50 NUCC-0202698

>50 ** NUCC-0202699

>50 >50 NUCC-0202700

>50 *** NUCC-0202701

>50 >50 NUCC-0202702

>50 >50 NUCC-0202703

>50 *** NUCC-0202704

>50 *** NUCC-0202705

>50 *** NUCC-0202706

>50 *** NUCC-0202707

>50 >50 NUCC-0202708

>50 >50 NUCC-0202709

>50 ** NUCC-0202710

>50 >50 NUCC-0202711

>50 ** NUCC-0202712

>50 * NUCC-0202713

>50 ** NUCC-0202714

>50 >50 NUCC-0202715

>50 ** NUCC-0202716

>50 >50 NUCC-0202717

>50 ** NUCC-0202718

>50 >50 NUCC-0202719

>50 >50 NUCC-0202720

>50 ** NUCC-0202721

>50 >50 NUCC-0202722

>50 >50 NUCC-0202723

>50 ** NUCC-0202724

>50 ** NUCC-0202725

>50 * NUCC-0202838

NT NUCC-0202839

NT NUCC-0202840

NT NUCC-0202841

NT NUCC-0202842

NT NUCC-0202843

NT

Example 4

Additional compounds that may be synthesized utilizing the methodsdisclosed herein may include the following compounds.

Example 5

Reference is made to U.S. Publication No. 2019/0062281, published onFeb. 28, 2019, the content of which is incorporated herein by referencein its entirety.

Example 6

The following additional compounds were synthesized utilizing themethods disclosed herein.

1H NMR: DMSO-d6 400 MHz: δ 10.13 (s, 1H), 7.34 (d, J=8.8 Hz, 1H), 7.22(d, J=7.6 Hz, 1H), 7.04-6.96 (m, 3H), 6.90 (dd, J=8.0, 1.6 Hz, 1H), 6.78(dd, J=8.8, 2.8 Hz, 1H), 6.68 (s, 1H), 4.32 (d, J=6.0 Hz, 2H), 3.73 (s,3H).

(M+1)=450.3

1H NMR: DMSO-d6 400 MHz: δ 10.01 (s, 1H), 7.77 (d, J=8.0 Hz, 1H),7.34-7.31 (m, 2H), 7.00 (d, J=2.8 Hz, 1H), 6.96-6.91 (m, 2H), 6.87 (dd,J=8.0, 1.2 Hz, 1H), 6.77 (dd, J=8.8, 2.8 Hz, 1H), 4.28 (d, J=6.0 Hz,2H), 4.01 (s, 3H).

(M+1)=450.2

1H NMR: DMSO-d6 400 MHz: δ 7.62 (d, J=8.0 Hz, 1H), 7.42 (d, J=9.2 Hz,1H), 7.02 (s, 1H), 6.99 (d, J=2.8 Hz, 1H), 6.94 (dd, J=9.2, 2.8 Hz, 1H),6.79 (s, 1H), 6.74 (dd, J=8.0, 1.2 Hz, 1H), 4.62 (s, 2H), 3.10 (s, 3H).

(M+1)=450.2

1H NMR: DMSO-d6 400 MHz: δ 10.25 (s, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.21(d, J=8.0 Hz, 1H), 7.01 (d, J=2.8 Hz, 1H), 6.95 (dd, J=9.2, 3.2 Hz, 1H),6.81 (s, 1H), 6.74 (d, J=8.0 Hz, 1H), 6.68 (s, 1H), 4.65 (s, 2H), 3.72(s, 3H), 3.11 (s, 3H).

(M+1)=464.4

1H NMR: DMSO-d6 400 MHz: δ 10.70 (s, 1H), 10.61 (s, 1H), 8.37 (d, J=2.4Hz, 1H), 8.11 (dd, J=8.8, 2.4 Hz, 1H), 7.73 (d, J=8.8 Hz, 1H), 7.54 (d,J=8.4 Hz, 2H), 7.46 (d, J=8.0 Hz, 1H), 6.81 (s, 1H), 3.79 (s, 3H).

(M+1)=464.1

1H NMR: DMSO-d6 400 MHz: δ 10.65 (s, 1H), 10.53 (s, 1H), 8.37 (d, J=2.4Hz, 1H), 8.12 (dd, J=8.8, 2.4 Hz, 1H), 8.00 (d, J=8.0 Hz, 1H), 7.32 (d,J=8.8 Hz, 1H), 7.56 (d, J=2.0 Hz, 1H), 7.52 (dd, J=8.4, 1.6 Hz, 1H),7.46 (s, 1H), 4.06 (s, 3H).

(M+1)=464.1

1H NMR: DMSO-d6 400 MHz: δ 13.76 (s, 1H), 9.24 (s, 1H), 7.76 (d, J=8.4Hz, 2H), 7.65 (dd, J=8.4, 2.0 Hz, 1H), 7.40-7.37 (m, 2H), 7.31-7.27 (m,2H), 7.02 (s, 1H), 6.77 (s, 1H), 5.15 (s, 2H).

(M+1)=581.1

1H NMR: DMSO-d6 400 MHz: δ 7.85 (d, J=1.6 Hz, 1H), 7.73 (d, J=8.4 Hz,1H), 7.69 (dd, J=8.4, 2.0 Hz, 1H), 7.41-7.37 (m, 2H), 7.32-7.30 (m, 2H),6.97 (s, 1H), 6.73 (s, 1H), 5.13 (s, 2H), 3.69 (s, 3H).

(M+1)=597.1

1H NMR: DMSO-d6 400 MHz: δ 9.55 (s, 1H), 7.80 (d, J=2.0 Hz, 1H), 7.74(d, J=8.0 Hz, 1H), 7.69 (dd, J=8.0, 1.6 Hz, 1H), 7.40-7.37 (m, 2H),7.32-7.30 (m, 2H), 7.14 (s, 1H), 6.99 (s, 1H), 5.14 (s, 2H), 4.02 (s,3H).

(M+1)=597.1

1H NMR: DMSO-d6 400 MHz: δ 7.78 (d, J=2.4 Hz, 1H), 7.63 (d, J=8.8 Hz,1H), 7.58 (d, J=8.4 Hz, 1H), 7.49 (dd, J=8.8, 2.4 Hz, 1H), 7.11 (s, 1H),6.98 (d, J=1.6 Hz, 1H), 6.78 (dd, J=8.0, 2.4 Hz, 1H), 3.39 (s, 3H).

(M+1)=464.3

1H NMR: DMSO-d6 400 MHz: δ 10.38 (s, 1H), 7.68 (d, J=8.8 Hz, 2H), 7.58(dd, J=8.4, 2.0 Hz, 1H), 7.17 (d, J=7.6 Hz, 1H), 6.97 (s, 1H), 6.79 (d,J=7.6 Hz, 1H), 6.69 (s, 1H), 3.67 (s, 3H), 3.41 (s, 3H).

(M+1)=478.3

1H NMR: EW14114-319-P2D DMSO-d6 400 MHz δ 10.29 (s, 1H), 7.75 (d, J=2.4Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.63 (d, J=8.8 Hz, 1H), 7.50 (dd,J=8.8, 2.4 Hz, 1H), 7.33 (s, 1H), 6.95 (d, J=1.2 Hz, 1H), 6.76 (dd,J=8.0, 1.4 Hz, 1H), 4.01 (s, 3H), 3.40 (s, 3H)

(M+1)=478.3

1H NMR: DMSO-d6 400 MHz: δ 10.64 (s, 1H), 8.37 (d, J=2.4 Hz, 1H), 8.11(dd, J=8.8, 2.4 Hz, 1H) 7.88 (d, J=8.0 Hz, 1H), 7.72 (d, J=8.8 Hz, 1H),7.56-7.53 (m, 2H), 7.23 (s, 1H).

(M+1)=450.1

1H NMR: DMSO-d6 400 MHz: δ 7.63 (d, J=8.0 Hz, 1H), 7.32 (d, J=9.6 Hz,1H), 7.03 (s, 1H), 7.00 (d, J=2.8 Hz, 1H), 6.99-6.95 (m, 2H), 6.88 (dd,J=8.0, 0.8 Hz, 1H), 6.76 (dd, J=8.8, 2.8 Hz, 1H), 4.29 (d, J=6.4 Hz,1H).

(M+1)=436.3

1H NMR (500 MHz, Chloroform-d) δ 7.95 (br s, 1H), 7.50 (d, J=8.2 Hz,1H), 7.43 (s, 1H), 7.28 (d, J=8.2 Hz, 1H), 7.07 (d, J=8.1 Hz, 1H), 6.84(s, 1H), 6.63 (dd, J=7.9, 1.9 Hz, 1H), 6.47 (s, 1H), 5.12 (t, J=5.8 Hz,1H), 4.02 (t, J=6.1 Hz, 2H), 3.69 (s, 3H), 3.45 (q, J=6.1 Hz, 2H), 1.40(s, 9H).

(M+1)=550.4

1H NMR (500 MHz, Chloroform-d) δ 7.56 (d, J=1.5 Hz, 1H), 7.43 (d, J=8.2Hz, 1H), 7.35 (d, J=8.2 Hz, 1H), 7.24-7.19 (m, 1H), 6.89 (s, 1H), 6.77(d, J=2.2 Hz, 1H), 6.61-6.51 (m, 1H), 4.99 (s, 1H), 4.10-3.95 (m, 5H),3.43 (d, J=6.0 Hz, 2H), 1.41 (s, 10H).

(M+1)=550.4

1H NMR (500 MHz, Chloroform-d) δ 7.54 (d, J=8.1 Hz, 1H), 7.44 (d, J=1.5Hz, 1H), 7.30 (d, J=8.1 Hz, 1H), 7.12 (d, J=8.1 Hz, 1H), 6.74 (d, J=2.0Hz, 1H), 6.68 (dd, J=8.1, 2.0 Hz, 1H), 6.56 (s, 1H), 6.52 (s, 1H), 3.90(t, J=7.7 Hz, 2H), 3.72 (s, 3H), 1.64 (td, J=7.3, 6.6, 2.5 Hz, 2H),1.44-1.16 (m, 6H), 0.94-0.77 (m, 3H).

1H NMR (500 MHz, Methanol-d4) δ 7.75-7.60 (m, 2H), 7.49 (dd, J=8.0, 1.5Hz, 1H), 7.22 (d, J=8.7 Hz, 1H), 6.86 (dq, J=4.4, 2.1 Hz, 2H), 6.53 (s,1H), 4.29 (t, J=6.1 Hz, 2H), 3.70 (s, 3H), 3.49 (q, J=7.0 Hz, 1H), 3.25(t, J=6.1 Hz, 2H), 1.18 (t, J=7.0 Hz, 3H).

1H NMR (500 MHz, Methanol-d4) δ 7.75-7.67 (m, 2H), 7.63 (d, J=8.2 Hz,1H), 7.44 (d, J=8.1 Hz, 1H), 7.24 (s, 1H), 6.88 (d, J=2.1 Hz, 1H), 6.79(dd, J=8.3, 2.2 Hz, 1H), 4.26 (t, J=6.1 Hz, 2H), 4.04 (s, 3H), 3.49 (qd,J=7.1, 1.0 Hz, 1H), 3.22 (t, J=6.1 Hz, 2H), 1.38-1.23 (m, 4H), 1.18 (td,J=7.0, 1.0 Hz, 2H), 0.97-0.78 (m, 2H).

1H NMR (500 MHz, Methanol-d4) δ 7.70 (d, J=8.1 Hz, 1H), 7.57 (s, 1H),7.45 (d, J=8.1 Hz, 1H), 7.21 (d, J=8.1 Hz, 1H), 6.82 (dd, J=8.1, 2.1 Hz,1H), 6.74 (d, J=2.2 Hz, 1H), 6.56 (s, 1H), 3.94 (t, J=7.6 Hz, 2H), 3.71(s, 3H), 3.37 (s, 1H), 1.71 (q, J=7.5 Hz, 2H), 1.01 (t, J=7.4 Hz, 3H).

1H NMR (500 MHz, Methanol-d4) δ 7.70 (d, J=8.1 Hz, 1H), 7.57 (s, 1H),7.45 (d, J=8.1 Hz, 1H), 7.21 (d, J=8.1 Hz, 1H), 6.81 (dd, J=8.1, 2.0 Hz,1H), 6.74 (d, J=2.1 Hz, 1H), 6.56 (s, 1H), 3.98 (dd, J=9.1, 6.0 Hz, 2H),3.71 (s, 3H), 1.67 (ddt, J=9.1, 7.7, 3.6 Hz, 2H), 1.43 (q, J=7.5 Hz,2H), 0.98 (t, J=7.4 Hz, 4H).

1H NMR: MeOD 400 MHz: δ 7.78-7.68 (m, 3H), 7.25-7.23 (m, 1H), 6.74-6.72(m, 1H), 6.60 (s, 1H), 3.81 (s, 3H), 3.52 (s, 2H), 1.91 (s, 3H),1.76-1.60 (m, 6H), 1.47 (s, 6H).

1H NMR: CDCL3 400 MHz: δ 7.85 (s, 1H), 7.84-7.62 (m, 2H), 7.32-7.22 (m,10H), 7.22-7.09 (m, 1H), 6.71-6.69 (m, 1H), 6.55 (s, 1H), 6.26 (s, 1H),5.09 (s, 1H), 3.82 (s, 3H).

1H NMR: DMSO-d6 400 MHz: δ 8.97 (s, 1H), 7.80-7.78 (m, 2H), 7.71-7.68(m, 1H), 7.18-7.08 (m, 56H), 6.71-6.68 (m, 2H), 5.50-5.49 (m, 1H), 3.72(s, 3H), 2.40 (d, 2H, J=7.2 Hz), 1.84-1.74 (m, 1H), 1.38 (d, 2H, J=6.4Hz), 0.83 (d, 6H, J=6.8 Hz).

Inhibitory Activity of NUCC Compounds Molecule Name CMV Reporter Ebocreporter NUCC-0202980 * * NUCC-0202981 — * NUCC-0202982 — **NUCC-0202983 * ** NUCC-0202984 * *** NUCC-0202985 — — NUCC-0202986 * ***NUCC-0202987 — *** NUCC-0202988 — — NUCC-0202989 * * NUCC-0202990 — *NUCC-0202991 — * NUCC-0202992 * ** NUCC-0202993 * ** NUCC-0226158 NT NTNUCC-0226159 NT NT NUCC-0226160 NT NT NUCC-0226161 NT NT NUCC-0226162 NTNT NUCC-0226176 NT NT NUCC-0226177 NT NT NUCC-0226212 NT NT NUCC-0226217NT NT NUCC-0226218 NT NT IC50: *** <5 uM; ** 5-10 uM: * 10-50 uM; — >50uM; NT = not tested

In the foregoing description, it will be readily apparent to one skilledin the art that varying substitutions and modifications may be made tothe invention disclosed herein without departing from the scope andspirit of the invention. The invention illustratively described hereinsuitably may be practiced in the absence of any element or elements,limitation or limitations which is not specifically disclosed herein.The terms and expressions which have been employed are used as terms ofdescription and not of limitation, and there is no intention that in theuse of such terms and expressions of excluding any equivalents of thefeatures shown and described or portions thereof, but it is recognizedthat various modifications are possible within the scope of theinvention. Thus, it should be understood that although the presentinvention has been illustrated by specific embodiments and optionalfeatures, modification and/or variation of the concepts herein disclosedmay be resorted to by those skilled in the art, and that suchmodifications and variations are considered to be within the scope ofthis invention.

Citations to a number of patent and non-patent references may be madeherein. The cited references are incorporated by reference herein intheir entireties. In the event that there is an inconsistency between adefinition of a term in the specification as compared to a definition ofthe term in a cited reference, the term should be interpreted based onthe definition in the specification.

We claim:
 1. A compound having a formula I:

wherein R¹ is hydrogen or R¹ is aryl, alkylaryl, alkyldiaryl,heteroaryl, alkylheteroaryl, cycloalkyl, or cycloheteroalkyl, optionallyR¹ is substituted at one or more positions with one or more of alkyl,alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano,carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl; Alk¹is straight-chain or branched alkylenyl; n is 0 or 1; X is O, NH, N(R⁸),N(R⁸)CH(O), or CH(O)N(R⁸); p is 0 or 1; Alk² is straight-chain orbranched alkylenyl; q is 0 or 1; Y is C, CH, C-halo, C-haloalkyl, or N;Z is C, CH, C-halo, C-haloalkyl, or N; m is 0 or 1; R² is hydrogen,halo, or R² is alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl, orcycloheteroalkyl, optionally R² is substituted at one or more positionswith one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; R³ is hydrogen, alkyl, alkoxy, haloalkyl, haloalkoxy,alkenyl, aryl, alkylaryl, hydroxyl, halo, carboxyamido optionallysubstituted with alkyl, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; R⁴ is present or absent and when present R⁴ is hydrogen,amino, alkyl, or R⁴ is aryl or alkylaryl; R⁴ optionally is substitutedat one or more positions with one or more of alkyl, alkoxy, haloalkyl,haloalkoxy, aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, alkoxycarbonyl, aryloxy, and alkylaryloxy; W is C orN; R⁵ is present or absent and when present R⁵ is hydrogen, alkyl,alkoxy, haloalkyl, haloalkoxy, hydroxyl, or halo; R⁶ is present orabsent and when present R⁶ is hydrogen, amino, alkyl, or R⁶ is aryl oralkylaryl; R⁶ optionally is substituted at one or more positions withone or more of alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl,halo, cyano, amido, hydrazonyl, carbonyl, carboxyl, alkoxycarbonyl,aryloxy, and alkylaryloxy, or R⁶ and R⁵ together form a ring structurehaving a formula

r is 0 or 1; R⁷ is hydrogen, halo, or R⁷ is alkyl, aryl, alkylaryl,heteroaryl, cycloalkyl, or cycloheteroalkyl, optionally R⁷ issubstituted at one or more positions with one or more of alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, alkoxycarbonyl, oxoaryl, and oxoheteroaryl; and R⁸is hydrogen, alkyl, aryl, or alkylaryl optionally substituted with halo;with the proviso that at least one of R⁴ and R⁶ is absent; optionallywith the proviso that if R⁵ is hydrogen, then p is 1 and m is 1;optionally with the proviso that if R¹(Alk¹)_(n)(X)_(p)(Alk²)_(q)- ishydrogen, hydroxyl, or alkyl, and R⁵ is hydroxyl, then m is 1, or atleast one of R² and R³ is not hydrogen; and optionally with the provisothat no more than two of W, Y, and Z are N.
 2. The compound of claim 1,wherein at least one of R² and R⁷ is aryl, alkylaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, and R² and R⁷ optionally aresubstituted at one or more positions with one or more of alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, and alkoxycarbonyl.
 3. The compound of claim 1,wherein m is 0 and R² is hydrogen; or wherein R⁷ is hydrogen.
 4. Thecompound of claim 1, wherein R² is aryl, alkylaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, and R² optionally is substituted at oneor more positions with one or more of alkyl, alkoxy, haloalkyl,haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, and alkoxycarbonyl; and wherein R⁷ is hydrogen.
 5. Thecompound of claim 1, wherein m is 0 and R² is hydrogen; and wherein R⁷is aryl, alkylaryl, heteroaryl, cycloalkyl, or cycloheteroalkyl, and R⁷optionally is substituted at one or more positions with one or more ofalkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano,carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl.
 6. Thecompound of claim 1 having a formula I(i) or I(ii):


7. The compound of claim 1 having a formula selected from Ia(i), Ia(ii),Ib(i), Ib(ii), Ic(i), and Ic(ii):


8. A compound having a formula II:

wherein R¹ is hydrogen or R¹ is aryl, alkylaryl, alkyldiaryl,heteroaryl, alkylheteroaryl, cycloalkyl, or cycloheteroalkyl, optionallyR¹ is substituted at one or more positions with one or more of alkyl,alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano,carboxyamido, hydrazonyl, carbonyl, carboxyl, and alkoxycarbonyl; Alk¹is straight-chain or branched alkylenyl; n is 0 or 1; X is O, NH, N(R⁸),N(R⁸)CH(O), or CH(O)N(R⁸); p is 0 or 1; Alk² is straight-chain orbranched alkylenyl; q is 0 or 1; Y is C, CH, or N; m is 0 or 1; R² ishydrogen, halo, or R² is alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl,or cycloheteroalkyl, optionally R² is substituted at one or morepositions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; R³ is hydrogen, alkyl, alkoxy, haloalkyl, haloalkoxy,alkenyl, aryl, alkylaryl, hydroxyl, halo, carboxyamido optionallysubstituted with alkyl, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; R⁴ is present or absent and when present R⁴ is hydrogen,amino, alkyl, or R⁴ is aryl or alkylaryl; R⁴ optionally is substitutedat one or more positions with one or more of alkyl, alkoxy, haloalkyl,haloalkoxy, aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, alkoxycarbonyl, aryloxy, and alkylaryloxy; R⁶ ispresent or absent and when present R⁶ is hydrogen, amino, alkyl, or R⁶is aryl or alkylaryl; R⁶ optionally is substituted at one or morepositions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, amido, hydrazonyl, carbonyl, carboxyl,alkoxycarbonyl, aryloxy, and alkylaryloxy; r is 0 or 1; R⁷ is hydrogen,halo, or R⁷ is alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl, orcycloheteroalkyl, optionally R⁷ is substituted at one or more positionswith one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl,alkoxycarbonyl, oxoaryl, and oxoheteroaryl; and R⁸ is hydrogen, alkyl,aryl, or alkylaryl optionally substituted with halo; with the provisothat at least one of R⁴ and R⁶ is absent.
 9. The compound of claim 4having a formula IIa:


10. A compound having a formula III:

wherein: R¹ is hydrogen or aryl, alkylaryl, alkyldiaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, optionally R¹ is substituted at one ormore positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, and alkoxycarbonyl; Alk¹ is straight-chain or branchedalkylenyl (e.g., a diradical of a straight-chain or branched C₁-C₆ alkylgroup); n is 0 or 1; X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸); p is0 or 1; Alk² is straight-chain or branched alkylenyl (e.g., a diradicalof a straight-chain or branched C₁-C₆ alkyl group); q is 0 or 1; m is 0or 1; R² is hydrogen or halo, or R² is alkyl, aryl, alkylaryl,heteroaryl, cycloalkyl, or cycloheteroalkyl, optionally R² issubstituted at one or more positions with one or more of alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, and alkoxycarbonyl; R⁴ is hydrogen, amino, alkyl,aryl, or alkylaryl; R⁴ optionally is substituted at one or morepositions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, alkoxycarbonyl, aryloxy, and alkylaryloxy; R⁵ is hydrogen,alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, or halo; r is 0 or 1; R⁷is hydrogen or halo, or R⁷ is alkyl, aryl, alkylaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, optionally R⁷ is substituted at one ormore positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; and R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionallysubstituted with halo.
 11. The compound of claim 10 having a formulaIIIa or IIb:


12. A compound having a formula IV:

wherein: R¹ is hydrogen, aryl, alkylaryl, alkyldiaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, optionally R¹ is substituted at one ormore positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, and alkoxycarbonyl; Alk¹ is straight-chain or branchedalkylenyl (e.g., a diradical of a straight-chain or branched C₁-C₆ alkylgroup); n is 0 or 1; X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸); p is0 or 1; Alk² is straight-chain or branched alkylenyl; g is 0 or 1; Y isC or N; m is 0 or 1; R² is hydrogen or halo, or R² is alkyl, aryl,alkylaryl, heteroaryl, cycloalkyl, or cycloheteroalkyl, optionally R² issubstituted at one or more positions with one or more of alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, and alkoxycarbonyl; R⁴ is hydrogen, amino, alkyl,aryl, or alkylaryl; R⁴ optionally is substituted at one or morepositions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, alkoxycarbonyl, aryloxy, and alkylaryloxy; W is C or N; R⁵ ispresent or absent and when present R⁵ is hydrogen, alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, or halo; r is 0 or 1; R⁷ is hydrogen orhalo, or R⁷ is alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl, orcycloheteroalkyl, optionally R⁷ is substituted at one or more positionswith one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; and R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionallysubstituted with halo.
 13. A compound having a formula V:

wherein: R¹ is hydrogen or aryl, alkylaryl, alkyldiaryl, heteroaryl,cycloalkyl, or cycloheteroalkyl, optionally R¹ is substituted at one ormore positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy,aryl, hydroxyl, halo, cyano, carboxyamido, hydrazonyl, carbonyl,carboxyl, and alkoxycarbonyl; Alk¹ is straight-chain or branchedalkylenyl (e.g., a diradical of a straight-chain or branched C₁-C₆ alkylgroup); n is 0 or 1; X is O, NH, N(R⁸), N(R⁸)CH(O), or CH(O)N(R⁸); p is0 or 1; Alk² is straight-chain or branched alkylenyl (e.g., a diradicalof a straight-chain or branched C₁-C₆ alkyl group); g is 0 or 1; m is 0or 1; R² is hydrogen or halo, or R² is alkyl, aryl, alkylaryl,heteroaryl, cycloalkyl, or cycloheteroalkyl, optionally R² issubstituted at one or more positions with one or more of alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxyamido, hydrazonyl,carbonyl, carboxyl, and alkoxycarbonyl; R⁵ is hydrogen, alkyl, alkoxy,haloalkyl, haloalkoxy, hydroxyl, or halo; r is 0 or 1; R⁷ is hydrogen orhalo, or R⁷ is alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl, orcycloheteroalkyl, optionally R⁷ is substituted at one or more positionswith one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl,halo, cyano, carboxyamido, hydrazonyl, carbonyl, carboxyl, andalkoxycarbonyl; and R⁸ is hydrogen, alkyl, aryl, or alkylaryl optionallysubstituted with halo.
 14. A pharmaceutical composition comprising acompound of claim 1 and a suitable pharmaceutical carrier, excipient, ordiluent.
 15. A method of treating cancer comprising administering thecomposition of claim 14 to a subject having cancer.
 16. A pharmaceuticalcomposition comprising a compound of claim 8 and a suitablepharmaceutical carrier, excipient, or diluent.
 17. A method of treatingcancer comprising administering the composition of claim 16 to a subjecthaving cancer.
 18. A pharmaceutical composition comprising a compound ofclaim 10 and a suitable pharmaceutical carrier, excipient, or diluent.19. A method of treating cancer comprising administering the compositionof claim 18 to a subject having cancer.
 20. A pharmaceutical compositioncomprising a compound of claim 12 and a suitable pharmaceutical carrier,excipient, or diluent.
 21. A method of treating cancer comprisingadministering the composition of claim 20 to a subject having cancer.22. A pharmaceutical composition comprising a compound of claim 13 and asuitable pharmaceutical carrier, excipient, or diluent.
 23. A method oftreating cancer comprising administering the composition of claim 22 toa subject having cancer.